Lipid compositions containing bioactive fatty acids

ABSTRACT

Provided herein is technology relating to lipid compositions containing bioactive fatty acids and particularly, but not exclusively, to compositions and methods related to the production and use of structured lipid compositions containing sciadonic and/or pinoleic acid alone or in combination with other bioactive fatty acids including, but not limited to, eicosapentaenoic acid, docosahexaenoic acid, conjugated linoleic acid, and non-β-oxidizable fatty acid analogues such as tetradecylthioacetic acid.

CROSS-REFERENCE TO RELATED APPLICATIONS

This application is a continuation of U.S. patent application Ser. No.14/774,432, filed Sep. 10, 2015, allowed as U.S. Pat. No. 10,154,979,which is a 371 U.S. National Phase Entry of International ApplicationNo. PCT/US2014/022553, filed Mar. 10, 2014, which claims priority toU.S. Provisional Patent Application No. 61/775,836, filed Mar. 11, 2013,the contents of which are incorporated by reference in their entireties.

FIELD OF TECHNOLOGY

Provided herein is technology relating to lipid compositions containingbioactive fatty acids and particularly, but not exclusively, tocompositions and methods related to the production and use of structuredlipid compositions containing sciadonic and/or pinoleic acid alone or incombination with other bioactive fatty acids including, but not limitedto, eicosapentaenoic acid, docosahexaenoic acid, conjugated linoleicacid, and non-β-oxidizable fatty acid analogues such astetradecylthioacetic acid.

BACKGROUND

Bioactive fatty acids have been implicated for the treatment of variousdiseases and conditions. Bioactive fatty acids from natural sources haveformed the basis for many popular and successful dietary supplementsincluding various fish oils.

A number of bioactive fatty acids from a variety of sources have beenidentified including sciadonic acid, pinolenic acid, eicosapentaenoicacid, docosahexaenoic acid, and conjugated linoleic acid, just to name afew. Additionally, non-β-oxidizable fatty acid analogues such astetradecylthioacetic acid have been shown to have excellent bioactivity.However, the efficacy of bioactive fatty acids in treating variousdiseases and conditions has been disputed.

What is needed in the art are improved compounds, compositions andformulations that enhance the usefulness of bioactive fatty acid fortreating particular diseases and conditions.

SUMMARY

Provided herein is technology relating to lipid compositions containingbioactive fatty acids and particularly, but not exclusively, tocompositions and methods related to the production and use of structuredlipid compositions containing sciadonic and/or pinoleic acid alone or incombination with other bioactive fatty acids including, but not limitedto, eicosapentaenoic acid, docosahexaenoic acid, conjugated linoleicacid, and non-β-oxidizable fatty acid analogues such astetradecylthioacetic acid.

In some embodiments, the present invention provides a bioactive lipidcomposition comprising:

a first lipid component comprising at least onenon-methylene-interrupted fatty acid moiety and a second lipid componentcomprising at least one bioactive fatty acid moiety selected from thegroup consisting of an omega-3 fatty acid moiety, a non-beta-oxidizablefatty acid moiety, and a conjugated linoleic acid moiety wherein thebioactive composition comprises at least 1% of the first lipid componentw/w and at least 1% of the second lipid component w/w.

In some embodiments, the non-methylene-interrupted fatty acid moiety isselected from the group consisting of a 5,11,14-eicosatrienoic acidmoiety, a 5,9,12-cis-octadecatrienoic acid moiety; and a5,11,14,17-eicosatetraenoic acid moiety and combinations thereof. Insome embodiments, the omega-3 fatty acid moiety is selected from thegroup consisting of an all-cis-5,8,11,14,17-eicosapentaenoic acidmoiety, an all-cis-7,10,13,16,19-docosapentaenoic acid moiety, and anall-cis-4,7,10,13,16,19-docosahexaenoic acid moiety and combinationsthereof. In some embodiments, the non-beta-oxidizable fatty acid moietyis selected from the group consisting of a tetradecylthioacetic acid(TTA) moiety and a tetradecylselenoacetic acid (TSA) moiety andcombinations thereof. In some embodiments, the conjugated linoleic acidmoiety is selected from the group consisting of a c9,t11 conjugatedlinoleic acid moiety, a t10,c12 conjugated linoleic acid moiety, at9,t11 conjugated linoleic acid moiety, a t10,t12 conjugated linoleicacid moiety and combinations thereof.

In some embodiments, the at least one non-methylene-interrupted fattyacid moiety is selected from the group consisting free fatty acids,acylglycerides, phospholipids and esters comprising the at least onenon-methylene-interrupted fatty acid moiety. In some embodiments, the atleast one omega-3 fatty acid moiety is selected from the groupconsisting free fatty acids, acylglycerides, phospholipids and esterscomprising the at least one omega-3 fatty acid moiety. In someembodiments, the at least one non-beta-oxidizable fatty acid moiety isselected from the group consisting free fatty acids, acylglycerides,phospholipids and esters comprising the at least one non-beta-oxidizablefatty acid moiety. In some embodiments, the at least one conjugatedlinoleic acid moiety is selected from the group consisting free fattyacids, acylglycerides, phospholipids and esters comprising the at leastone conjugated linoleic acid moiety.

In some embodiments, the composition comprises at least 5%, 10%, 20%,30%, 40%, 50%, 60%, 70%, 80%, 90%, 95%, or 99% w/w of the first lipidcomponent. In some embodiments, the composition comprises at least 5%,10%, 20%, 30%, 40%, 50%, 60%, 70%, 80%, 90%, 95%, or 99% w/w of thesecond lipid component.

In some embodiments, the compositions of the present invention comprisedefined ratios of bioactive fatty acids. The bioactive fatty acids arepreferably provided in the compositions as free fatty acids or asderivatives comprising a fatty acid moiety such as fatty acid esters(e.g., ethyl esters), acylglycerides (e.g., triglycerides), orphospholipids. The ratios refer to the weight ratios of the bioactivefatty acids in the compositions. Accordingly, in some embodiments, thecompositions comprise a ratio of non-methylene-interrupted fatty acid:one or more other bioactive fatty acids of from 1:50 to 1:1, from 1:20to 1:1, from 1:10 to 1:1, from 1:5 to 1:1, from 1:4 to 1:1, from 1:3 to1:1, from 1:2 to 1:1, or from 1.5:1 to 1:1. In some embodiments, thecompositions comprise a ratio of non-methylene-interrupted fattyacid:one or more other bioactive fatty acids of from 1:50 to 1:2, from1:20 to 1:2, from 1:10 to 1:2, from 1:5 to 1:2, from 1:4 to 1:2, or from1:3 to 1:2. In some embodiments, the compositions comprise a ratio ofnon-methylene-interrupted fatty acid:one or more other bioactive fattyacids of from 1:50 to 1:3, from 1:20 to 1:3, from 1:10 to 1:3, from 1:5to 1:3, or from 1:4 to 1:3. In some embodiments, the compositionscomprise a ratio of non-methylene-interrupted fatty acid:one or moreother bioactive fatty acids of from 1:50 to 1:5, from 1:20 to 1:5, orfrom 1:10 to 1:5. In some embodiments, the compositions comprise a ratioof non-methylene-interrupted fatty acid:one or more other bioactivefatty acids of from 1:50 to 1:10, from 1:40 to 1:10, from 1:30 to 1:10,or from 1:20 to 1:10. In some embodiments, the compositions comprise aratio of one or more bioactive fatty acids that are not anon-methylene-interrupted fatty acid:non-methylene-interrupted fattyacid of from 1:50 to 1:2, from 1:20 to 1:2, from 1:10 to 1:2, from 1:5to 1:2, from 1:4 to 1:2, or from 1:3 to 1:2. In some embodiments, thecompositions comprise a ratio of one or more bioactive fatty acids thatare not a non-methylene-interrupted fatty acid:non-methylene-interruptedfatty acid of from 1:50 to 1:3, from 1:20 to 1:3, from 1:10 to 1:3, from1:5 to 1:3, or from 1:4 to 1:3. In some embodiments, the compositionscomprise a ratio of one or more bioactive fatty acids that are not anon-methylene-interrupted fatty acid:non-methylene-interrupted fattyacid of from 1:50 to 1:5, from 1:20 to 1:5, or from 1:10 to 1:5. In someembodiments, the compositions comprise a ratio of one or more bioactivefatty acids that are not a non-methylene-interrupted fattyacid:non-methylene-interrupted fatty acid of from 1:50 to 1:10, from1:40 to 1:10, from 1:30 to 1:10, or from 1:20 to 1:10. In someembodiments, the one or more other bioactive fatty acid is one or moreomega-3 fatty acids. In some embodiments, the omega-3 fatty acid isdocosahexaenoic acid (DHA). In some embodiments, the omega-3 fatty acidis eicosapentaenoic acid (EPA). In some embodiments, the omega-3 fattyacid is docosapentaenoic acid (DPA). In some embodiments, the one ormore omega-3 fatty acids are a combination of DHA and EPA. In someembodiments, the one or more bioactive fatty acids is conjugatedlinoleic acid. In some embodiments, the one or more bioactive fattyacids are a combination of CLA and one or more omega-3 fatty acids,preferably DHA and EPA. In some embodiments, the one or more bioactivefatty acids is a non-beta-oxidizable fatty acid, preferablytetradecylthioacetic acid (TTA). In some embodiments, the one or morebioactive fatty acids are a combination of a non-beta-oxidizable fattyacid, preferably TTA, and one or more omega-3 fatty acids, preferablyDHA and EPA. In some embodiments, the composition further comprise atleast one pharmaceutically acceptable carrier. In some embodiments, thecomposition comprises an oil, powder, crystal, wax, emulsion, micelle,vesicle, or film.

In some embodiments, the present invention provides an oral deliveryvehicle, food product, nutritional supplement, dietary supplement orfunction food comprising the bioactive lipid composition of describedabove.

In some embodiments, the present invention provides a structuredphospholipid composition comprising phospholipid molecules of thefollowing structure:

wherein R1 and R2 are fatty acid moieties or —H and R3 is —H or aphospholipid head group such that the composition comprises at least 1%w/w of at least one non-methylene-interrupted fatty acid moiety and atleast 1% w/w of a second bioactive lipid moiety.

In some embodiments, the non-methylene-interrupted fatty acid moiety isselected from the group consisting of a 5,11,14-eicosatrienoic acidmoiety, a 5,9,12-cis-octadecatrienoic acid moiety; and a5,11,14,17-eicosatetraenoic acid moiety and combinations thereof. Insome embodiments, the second bioactive lipid moiety is selected from thegroup consisting of an omega-3 fatty acid moiety, a non-beta-oxidizablefatty acid moiety, a conjugated linoleic acid moiety and combinationsthereof. In some embodiments, the omega-3 fatty acid moiety is selectedfrom the group consisting of an all-cis-5,8,11,14,17-eicosapentaenoicacid moiety, an all-cis-7,10,13,16,19-docosapentaenoic acid moiety, andan all-cis-4,7,10,13,16,19-docosahexaenoic acid moiety and combinationsthereof. In some embodiments, the non-beta-oxidizable fatty acid moietyis selected from the group consisting of a tetradecylthioacetic acid(TTA) moiety and a tetradecylselenoacetic acid (TSA) moiety andcombinations thereof. In some embodiments, the conjugated linoleic acidmoiety is selected from the group consisting of a c9,t11 conjugatedlinoleic acid moiety, a t10,c12 conjugated linoleic acid moiety, at9,t11 conjugated linoleic acid moiety, a t10,t12 conjugated linoleicacid moiety and combinations thereof. In some embodiments, thecomposition comprises at least 5%, 10%, 20%, 30%, 40%, 50%, 60%, 70%,80%, 90%, 95%, or 99% w/w of the at least one non-methylene-interruptedfatty acid moiety. In some embodiments, the composition comprises atleast 5%, 10%, 20%, 30%, 40%, 50%, 60%, 70%, 80%, 90%, 95%, or 99% ofthe second bioactive lipid moiety.

In some embodiments, the phospholipids comprise a weight ratio ofnon-methylene-interrupted fatty acid:one or more other bioactive fattyacids of from 1:50 to 1:1, from 1:20 to 1:1, from 1:10 to 1:1, from 1:5to 1:1, from 1:4 to 1:1, from 1:3 to 1:1, from 1:2 to 1:1, or from 1.5:1to 1:1. In some embodiments, the compositions comprise a weight ratio ofnon-methylene-interrupted fatty acid:one or more other bioactive fattyacids of from 1:50 to 1:2, from 1:20 to 1:2, from 1:10 to 1:2, from 1:5to 1:2, from 1:4 to 1:2, or from 1:3 to 1:2. In some embodiments, thecompositions comprise a weight ratio of non-methylene-interrupted fattyacid:one or more other bioactive fatty acids of from 1:50 to 1:3, from1:20 to 1:3, from 1:10 to 1:3, from 1:5 to 1:3, or from 1:4 to 1:3. Insome embodiments, the compositions comprise a weight ratio ofnon-methylene-interrupted fatty acid:one or more other bioactive fattyacids of from 1:50 to 1:5, from 1:20 to 1:5, or from 1:10 to 1:5. Insome embodiments, the compositions comprise a weight ratio ofnon-methylene-interrupted fatty acid:one or more other bioactive fattyacids of from 1:50 to 1:10, from 1:40 to 1:10, from 1:30 to 1:10, orfrom 1:20 to 1:10. In some embodiments, the compositions comprise aweight ratio of one or more bioactive fatty acids that are not anon-methylene-interrupted fatty acid:non-methylene-interrupted fattyacid of from 1:50 to 1:2, from 1:20 to 1:2, from 1:10 to 1:2, from 1:5to 1:2, from 1:4 to 1:2, or from 1:3 to 1:2. In some embodiments, thecompositions comprise a weight ratio of one or more bioactive fattyacids that are not a non-methylene-interrupted fattyacid:non-methylene-interrupted fatty acid of from 1:50 to 1:3, from 1:20to 1:3, from 1:10 to 1:3, from 1:5 to 1:3, or from 1:4 to 1:3. In someembodiments, the compositions comprise a weight ratio of one or morebioactive fatty acids that are not a non-methylene-interrupted fattyacid:non-methylene-interrupted fatty acid of from 1:50 to 1:5, from 1:20to 1:5, or from 1:10 to 1:5. In some embodiments, the compositionscomprise a weight ratio of one or more bioactive fatty acids that arenot a non-methylene-interrupted fatty acid:non-methylene-interruptedfatty acid of from 1:50 to 1:10, from 1:40 to 1:10, from 1:30 to 1:10,or from 1:20 to 1:10. In some embodiments, the one or more otherbioactive fatty acid is one or more omega-3 fatty acids. In someembodiments, the omega-3 fatty acid is docosahexaenoic acid (DHA). Insome embodiments, the omega-3 fatty acid is eicosapentaenoic acid (EPA).In some embodiments, the omega-3 fatty acid is docosapentaenoic acid(DPA). In some embodiments, the one or more omega-3 fatty acids are acombination of DHA and EPA. In some embodiments, the one or morebioactive fatty acids is conjugated linoleic acid. In some embodiments,the one or more bioactive fatty acids are a combination of CLA and oneor more omega-3 fatty acids, preferably DHA and EPA. In someembodiments, the one or more bioactive fatty acids is anon-beta-oxidizable fatty acid, preferably tetradecylthioacetic acid(TTA). In some embodiments, the one or more bioactive fatty acids are acombination of a non-beta-oxidizable fatty acid, preferably TTA, and oneor more omega-3 fatty acids, preferably DHA and EPA.

In some embodiments, the compositions further comprise at least onepharmaceutically acceptable carrier. In some embodiments, thecomposition comprises an oil, powder, crystal, wax, emulsion, micelle,vesicle, or film.

In some embodiments, the present invention provides an oral deliveryvehicle, food product, nutritional supplement, dietary supplement orfunction food comprising the structured phospholipid compositiondescribed above.

In some embodiments, the present invention provides a structuredacylglycerol composition comprising acylglycerol molecules of thefollowing structure:

wherein R1, R2 and R3 are fatty acid moieties or —H such that thecomposition comprises at least 1% w/w of at least onenon-methylene-interrupted fatty acid moiety and at least 1% w/w of asecond bioactive lipid moiety. In some embodiments, thenon-methylene-interrupted fatty acid moiety is selected from the groupconsisting of a 5,11,14-eicosatrienoic acid moiety, a5,9,12-cis-octadecatrienoic acid moiety; and a5,11,14,17-eicosatetraenoic acid moiety and combinations thereof. Insome embodiments, the second bioactive lipid moiety is selected from thegroup consisting of an omega-3 fatty acid moiety, a non-beta-oxidizablefatty acid moiety, a conjugated linoleic acid moiety and combinationsthereof. In some embodiments, the omega-3 fatty acid moiety is selectedfrom the group consisting of an all-cis-5,8,11,14,17-eicosapentaenoicacid moiety, an all-cis-7,10,13,16,19-docosapentaenoic acid moiety, andan all-cis-4,7,10,13,16,19-docosahexaenoic acid moiety and combinationsthereof. In some embodiments, the non-beta-oxidizable fatty acid moietyis selected from the group consisting of a tetradecylthioacetic acid(TTA) moiety and a tetradecylselenoacetic acid (TSA) moiety andcombinations thereof. In some embodiments, the conjugated linoleic acidmoiety is selected from the group consisting of a c9,t11 conjugatedlinoleic acid moiety, a t10,c12 conjugated linoleic acid moiety, at9,t11 conjugated linoleic acid moiety, a t10,t12 conjugated linoleicacid moiety and combinations thereof. In some embodiments, thecomposition comprises at least 5%, 10%, 20%, 30%, 40%, 50%, 60%, 70%,80%, 90%, 95%, or 99% w/w of the at least one non-methylene-interruptedfatty acid moiety. In some embodiments, the composition comprises atleast 5%, 10%, 20%, 30%, 40%, 50%, 60%, 70%, 80%, 90%, 95%, or 99% ofthe second bioactive lipid moiety.

In some embodiments, the acylglycerols, preferably triglycerides,comprise a weight ratio of non-methylene-interrupted fatty acid:one ormore other bioactive fatty acids of from 1:50 to 1:1, from 1:20 to 1:1,from 1:10 to 1:1, from 1:5 to 1:1, from 1:4 to 1:1, from 1:3 to 1:1,from 1:2 to 1:1, or from 1.5:1 to 1:1. In some embodiments, thecompositions comprise a weight ratio of non-methylene-interrupted fattyacid:one or more other bioactive fatty acids of from 1:50 to 1:2, from1:20 to 1:2, from 1:10 to 1:2, from 1:5 to 1:2, from 1:4 to 1:2, or from1:3 to 1:2. In some embodiments, the compositions comprise a weightratio of non-methylene-interrupted fatty acid:one or more otherbioactive fatty acids of from 1:50 to 1:3, from 1:20 to 1:3, from 1:10to 1:3, from 1:5 to 1:3, or from 1:4 to 1:3. In some embodiments, thecompositions comprise a weight ratio of non-methylene-interrupted fattyacid:one or more other bioactive fatty acids of from 1:50 to 1:5, from1:20 to 1:5, or from 1:10 to 1:5. In some embodiments, the compositionscomprise a weight ratio of non-methylene-interrupted fatty acid:one ormore other bioactive fatty acids of from 1:50 to 1:10, from 1:40 to1:10, from 1:30 to 1:10, or from 1:20 to 1:10. In some embodiments, thecompositions comprise a weight ratio of one or more bioactive fattyacids that are not a non-methylene-interrupted fattyacid:non-methylene-interrupted fatty acid of from 1:50 to 1:2, from 1:20to 1:2, from 1:10 to 1:2, from 1:5 to 1:2, from 1:4 to 1:2, or from 1:3to 1:2. In some embodiments, the compositions comprise a weight ratio ofone or more bioactive fatty acids that are not anon-methylene-interrupted fatty acid:non-methylene-interrupted fattyacid of from 1:50 to 1:3, from 1:20 to 1:3, from 1:10 to 1:3, from 1:5to 1:3, or from 1:4 to 1:3. In some embodiments, the compositionscomprise a weight ratio of one or more bioactive fatty acids that arenot a non-methylene-interrupted fatty acid:non-methylene-interruptedfatty acid of from 1:50 to 1:5, from 1:20 to 1:5, or from 1:10 to 1:5.In some embodiments, the compositions comprise a weight ratio of one ormore bioactive fatty acids that are not a non-methylene-interruptedfatty acid:non-methylene-interrupted fatty acid of from 1:50 to 1:10,from 1:40 to 1:10, from 1:30 to 1:10, or from 1:20 to 1:10. In someembodiments, the one or more other bioactive fatty acid is one or moreomega-3 fatty acids. In some embodiments, the omega-3 fatty acid isdocosahexaenoic acid (DHA). In some embodiments, the omega-3 fatty acidis eicosapentaenoic acid (EPA). In some embodiments, the omega-3 fattyacid is docosapentaenoic acid (DPA). In some embodiments, the one ormore omega-3 fatty acids are a combination of DHA and EPA. In someembodiments, the one or more bioactive fatty acids is conjugatedlinoleic acid. In some embodiments, the one or more bioactive fattyacids are a combination of CLA and one or more omega-3 fatty acids,preferably DHA and EPA. In some embodiments, the one or more bioactivefatty acids is a non-beta-oxidizable fatty acid, preferablytetradecylthioacetic acid (TTA). In some embodiments, the one or morebioactive fatty acids are a combination of a non-beta-oxidizable fattyacid, preferably TTA, and one or more omega-3 fatty acids, preferablyDHA and EPA.

In some embodiments, the compositions further comprise at least onepharmaceutically acceptable carrier. In some embodiments, thecomposition comprises an oil, powder, crystal, wax, emulsion, micelle,vesicle, or film.

In some embodiments, the present invention provides an oral deliveryvehicle, food product, nutritional supplement, dietary supplement orfunction food comprising the structured acylglycerol compositiondescribed above.

In some embodiments, the present invention provides a bioactive lipidcomposition comprising at 1% w/w of the structured phospholipidcomposition as described above and at least 1% w/w of the structuredacylglyceride composition as described above.

In some embodiments, the present invention provides a method of treatinga subject comprising administering to the subject the bioactive lipidcomposition, structured phospholipid composition or structuredacylglyceride composition or oral delivery vehicle, food product,nutritional supplement, dietary supplement or function food as describedabove to a subject in need thereof. In some embodiments, theadministration or oral, topical, parenteral, enteral, transdermal,intradermal, intraocular, intravitreal, sublingual, or intravaginal.

In some embodiments, the present invention provides a method of reducingobesity, inducing weight loss, increasing lean body mass, increasingmuscularity, increasing muscle mass, improving body composition,alleviating one or more symptoms metabolic syndrome, treating diabetes,decreasing insulin resistance, reducing inflammation, improvingconcentration, memory, cognitive function, attention and treating,alleviating or improving one or more of the following diseases orconditions: restenosis, arteriosclerosis, coronary heart disease,thrombosis, myocardial infarction, stroke, hypertension, fatty liver,diabetes, hyperglycaemia, hyperinsulinemia, and stenosis, rheumatoidarthritis, systemic vasculitis, systemic lupus erythematosus, systemicsclerosis, dermatomyositis, polymyositis, various autoimmune endocrinedisorders (e.g. thyroiditis and adrenalitis), various immune mediatedneurological disorders (e.g. multiple sclerosis and myastenia gravis),various cardiovascular disorders (e.g. myocarditis, congestive heartfailure, arteriosclerosis and stable and unstable angina, and Wegenersgranulomatosis), inflammatory bowel diseases and colitis (e.g., Crohn'scolitis), nephritis, various inflammatory skin disorders (e.g.psoriasis, atopic dermatitis and food allergy) and acute and chronicallograft rejection after organ transplantation, comprising:administering to a subject in need thereof the bioactive lipidcomposition, structured phospholipid composition or structuredacylglyceride composition or oral delivery vehicle, food product,nutritional supplement, dietary supplement or function food of any ofclaims 1 to 37. In some embodiments, the administration or oral,topical, parenteral, enteral, transdermal, intradermal, intraocular,intravitreal, sublingual, or intravaginal.

In some embodiments, the present invention provides a method of treatingdiabetes, ameliorating the symptoms of diabetes, providing nutritionalsupport to a subject with diabetes, promoting healthy blood sugarlevels, supporting efficient insulin production and secretion, andsupporting healthy glucose metabolism, comprising: administering to asubject in need thereof the bioactive lipid composition, structuredphospholipid composition or structured acylglyceride composition or oraldelivery vehicle, food product, nutritional supplement, dietarysupplement or function food as described above. In some embodiments, theadministration or oral, topical, parenteral, enteral, transdermal,intradermal, intraocular, intravitreal, sublingual, or intravaginal.

Additional embodiments will be apparent to persons skilled in therelevant art based on the teachings contained herein.

DETAILED DESCRIPTION

Provided herein is technology relating to lipid compositions containingbioactive fatty acids and particularly, but not exclusively, tocompositions and methods related to the production and use of structuredlipid compositions containing sciadonic and/or pinoleic acid alone or incombination with other bioactive fatty acids including, but not limitedto, eicosapentaenoic acid, docosahexaenoic acid, conjugated linoleicacid, and non-β-oxidizable fatty acid analogues such astetradecylthioacetic acid.

This technology is described below, wherein the section headings are fororganizational purposes only and are not to be construed as limiting thedescribed subject matter in any way.

In this detailed description of the various embodiments, for purposes ofexplanation, numerous specific details are set forth to provide athorough understanding of the embodiments disclosed. One skilled in theart will appreciate, however, that these various embodiments may bepracticed with or without these specific details. In other instances,structures and devices are shown in block diagram form. Furthermore, oneskilled in the art can readily appreciate that the specific sequences inwhich methods are presented and performed are illustrative and it iscontemplated that the sequences can be varied and still remain withinthe spirit and scope of the various embodiments disclosed herein.

All literature and similar materials cited in this application,including but not limited to, patents, patent applications, articles,books, treatises, and internet web pages are expressly incorporated byreference in their entirety for any purpose. Unless defined otherwise,all technical and scientific terms used herein have the same meaning asis commonly understood by one of ordinary skill in the art to which thevarious embodiments described herein belongs. When definitions of termsin incorporated references appear to differ from the definitionsprovided in the present teachings, the definition provided in thepresent teachings shall control.

Definitions

To facilitate an understanding of the present technology, a number ofterms and phrases are defined below. Additional definitions are setforth throughout the detailed description.

Throughout the specification and claims, the following terms take themeanings explicitly associated herein, unless the context clearlydictates otherwise. The phrase “in one embodiment” as used herein doesnot necessarily refer to the same embodiment, though it may.Furthermore, the phrase “in another embodiment” as used herein does notnecessarily refer to a different embodiment, although it may. Thus, asdescribed below, various embodiments of the technology may be readilycombined, without departing from the scope or spirit of the technology.

In addition, as used herein, the term “or” is an inclusive “or” operatorand is equivalent to the term “and/or” unless the context clearlydictates otherwise. The term “based on” is not exclusive and allows forbeing based on additional factors not described, unless the contextclearly dictates otherwise. In addition, throughout the specification,the meaning of “a”, “an”, and “the” include plural references. Themeaning of “in” includes “in” and “on.”

As used herein, “feeding” refers to providing a substance, compound,composition, etc. to a living organism. For example, the substance,compound, composition, etc. may be an energy source, a carbon source, anutrient, or a source of other elements, molecules, and/or precursors ofbiological molecules that are used by the living organism and/or aremetabolized (e.g., catabolized, anabolized) by the living organism. Thesubstance, compound, composition, etc. is not necessarily a substance,compound, composition, etc. that the living organism encounters in itsnative milieu, but may be a synthetic substance, compound, composition,etc. or a natural substance, compound, composition, etc. that isnevertheless used by the living organism for metabolism. The substance,compound, composition, etc. may be added to a culture medium or asubstrate in which or on which the living organism lives and/or grows.

As used herein, “active” or “activity” refers to native or naturallyoccurring biological and/or immunological activity.

As used herein the term, “in vitro” refers to an artificial environmentand to processes or reactions that occur within an artificialenvironment. In vitro environments may include, but are not limited to,test tubes and cell cultures. The term “in vivo” refers to the naturalenvironment (e.g., an animal or a cell) and to processes or reactionsthat occur within a natural environment.

As used herein, the terms “subject” and “patient” refer to any animal,such as a mammal like a dog, cat, bird, livestock, and preferably ahuman (e g, a human with a disease such as obesity, diabetes, or insulinresistance).

As used herein, the term “individual” refers to vertebrates,particularly members of the mammalian species. The term includes but isnot limited to domestic animals, sports animals, primates, and humans.

As used herein, the term “effective amount” refers to the amount of acomposition sufficient to effect beneficial or desired results. Aneffective amount can be administered in one or more administrations,applications, or dosages and is not intended to be limited to aparticular formulation or administration route.

As used herein, the term “administration” refers to the act of giving adrug, prodrug, or other agent, or therapeutic treatment to a subject.Exemplary routes of administration to the human body can be through theeyes (ophthalmic), mouth (oral), skin (transdermal, topical), nose(nasal), lungs (inhalant), oral mucosa (buccal), ear, by injection(e.g., intravenously, subcutaneously, intratumorally, intraperitoneally,etc.), and the like.

As used herein, the term “co-administration” refers to theadministration of at least two agents or therapies to a subject. In someembodiments, the co-administration of two or more agents or therapies isconcurrent. In other embodiments, a first agent/therapy is administeredprior to a second agent/therapy. Those of skill in the art understandthat the formulations and/or routes of administration of the variousagents or therapies used may vary. The appropriate dosage forco-administration can be readily determined by one skilled in the art.In some embodiments, when agents or therapies are co-administered, therespective agents or therapies are administered at lower dosages thanappropriate for their administration alone. Thus, co-administration isespecially desirable in embodiments where the co-administration of theagents or therapies lowers the requisite dosage of a potentially harmful(e.g., toxic) agent.

As used herein, the term “pharmaceutical composition” refers to thecombination of an active agent with a carrier, inert or active, makingthe composition especially suitable for therapeutic use.

The terms “pharmaceutically acceptable” or “pharmacologicallyacceptable”, as used herein, refer to compositions that do notsubstantially produce adverse reactions, e.g., toxic, allergic, orimmunological reactions, when administered to a subject.

As used herein, the term “treating” includes reducing or alleviating atleast one adverse effect or symptom of a disease or disorder throughintroducing in any way a therapeutic composition of the presenttechnology into or onto the body of a subject. “Treatment” refers toboth therapeutic treatment and prophylactic or preventative measures,wherein the object is to prevent or slow down (lessen) the targetedpathologic condition or disorder. Those in need of treatment includethose already with the disorder as well as those prone to have thedisorder or those in whom the disorder is to be prevented.

As used herein, the term “sample” is used in its broadest sense. In onesense, it is meant to include a specimen or culture obtained from anysource, as well as biological and environmental samples.

Biological samples may be obtained from animals (including humans) andencompass fluids, solids, tissues, and gases. Biological samples includeblood products, such as plasma, serum and the like. Environmentalsamples include environmental material such as surface matter, soil,water, crystals and industrial samples. Such examples are not however tobe construed as limiting the sample types applicable to the presenttechnology.

As used herein, the terms “alkyl” and the prefix “alk-” are inclusive ofboth straight chain and branched chain saturated or unsaturated groups,and of cyclic groups, e.g., cycloalkyl and cycloalkenyl groups. Unlessotherwise specified, acyclic alkyl groups are from 1 to 6 carbons.Cyclic groups can be monocyclic or polycyclic and preferably have from 3to 8 ring carbon atoms. Exemplary cyclic groups include cyclopropyl,cyclopentyl, cyclohexyl, and adamantyl groups. Alkyl groups may besubstituted with one or more substituents or unsubstituted. Exemplarysubstituents include alkoxy, aryloxy, sulfhydryl, alkylthio, arylthio,halogen, alkylsilyl, hydroxyl, fluoroalkyl, perfluoralkyl, amino,aminoalkyl, disubstituted amino, quaternary amino, hydroxyalkyl,carboxyalkyl, and carboxyl groups. When the prefix “alk” is used, thenumber of carbons contained in the alkyl chain is given by the rangethat directly precedes this term, with the number of carbons containedin the remainder of the group that includes this prefix definedelsewhere herein. For example, the term “C₁-C₄ alkaryl” exemplifies anaryl group of from 6 to 18 carbons (e.g., see below) attached to analkyl group of from 1 to 4 carbons.

As used herein, the term “aryl” refers to a carbocyclic aromatic ring orring system. Unless otherwise specified, aryl groups are from 6 to 18carbons. Examples of aryl groups include phenyl, naphthyl, biphenyl,fluorenyl, and indenyl groups.

As used herein, the term “heteroaryl” refers to an aromatic ring or ringsystem that contains at least one ring heteroatom (e.g., O, S, Se, N, orP). Unless otherwise specified, heteroaryl groups are from 1 to 9carbons. Heteroaryl groups include furanyl, thienyl, pyrrolyl,imidazolyl, pyrazolyl, oxazolyl, isoxazolyl, thiazolyl, isothiazolyl,triazolyl, tetrazolyl, oxadiazolyl, oxatriazolyl, pyridyl, pyridazyl,pyrimidyl, pyrazyl, triazyl, benzofuranyl, isobenzofuranyl,benzothienyl, indole, indazolyl, indolizinyl, benzisoxazolyl,quinolinyl, isoquinolinyl, cinnolinyl, quinazolinyl, naphtyridinyl,phthalazinyl, phenanthrolinyl, purinyl, and carbazolyl groups.

As used herein, the term “heterocycle” refers to a non-aromatic ring orring system that contains at least one ring heteroatom (e.g., O, S, Se,N, or P). Unless otherwise specified, heterocyclic groups are from 2 to9 carbons. Heterocyclic groups include, for example, dihydropyrrolyl,tetrahydropyrrolyl, piperazinyl, pyranyl, dihydropyranyl,tetrahydropyranyl, dihydrofuranyl, tetrahydrofuranyl, dihydrothiophene,tetrahydrothiophene, and morpholinyl groups.

Aryl, heteroaryl, or heterocyclic groups may be unsubstituted orsubstituted by one or more substituents selected from the groupconsisting of C₁₋₆ alkyl, hydroxy, halo, nitro, C₁₋₆ alkoxy, C₁₋₆alkylthio, trifluoromethyl, C₁₋₆ acyl, arylcarbonyl, heteroarylcarbonyl,nitrile, C₁₋₆ alkoxycarbonyl, alkaryl (where the alkyl group has from 1to 4 carbon atoms), and alkheteroaryl (where the alkyl group has from 1to 4 carbon atoms).

As used herein, the term “alkoxy” refers to a chemical substituent ofthe formula —OR, where R is an alkyl group. By “aryloxy” is meant achemical substituent of the formula —OR′, where R′ is an aryl group.

As used herein, the term “C_(x-y) alkaryl” refers to a chemicalsubstituent of formula —RR′, where R is an alkyl group of x to y carbonsand R′ is an aryl group as defined elsewhere herein.

As used herein, the term “C_(x-y) alkheteraryl” refers to a chemicalsubstituent of formula RR″, where R is an alkyl group of x toy carbonsand R″ is a heteroaryl group as defined elsewhere herein.

As used herein, the term “halide” or “halogen” or “halo” refers tobromine, chlorine, iodine, or fluorine.

As used herein, the term “non-vicinal O, S, or N” refers to an oxygen,sulfur, or nitrogen heteroatom substituent in a linkage, where theheteroatom substituent does not form a bond to a saturated carbon thatis bonded to another heteroatom.

For structural representations where the chirality of a carbon has beenleft unspecified it is to be presumed by one skilled in the art thateither chiral form of that stereocenter is possible.

Embodiments of the Technology

Provided herein is technology relating to lipid compositions containingbioactive fatty acids and particularly, but not exclusively, tocompositions and methods related to the production and use of structuredlipid compositions containing sciadonic and/or pinoleic acid alone or incombination with other bioactive fatty acids including, but not limitedto, eicosapentaenoic acid, docosahexaenoic acid, conjugated linoleicacid, and non-β-oxidizable fatty acid analogues such astetradecylthioacetic acid. Below, sources of bioactive fatty acids,lipid compositions comprising bioactive fatty acids, methods for makingthe compositions and uses of the compositions are described.

Non-Methylene-Interrupted Fatty Acids

The term non-methylene-interrupted fatty acid, the acronym for which isNMIFA, refers to a fatty acid with a series of double bonds in which atleast one adjacent pair of double bonds is separated by at least twocarbon atoms, i.e., by a group other than a single methylene group.Examples of NMIFA include, but are not limited to,5,11,14-eicosatrienoic acid; 5,9,12-cis-octadecatrienoic acid; and5,11,14,17-eicosatetraenoic acid. Preferred NMIFAs have the followingformula, wherein the NMIFA is an acid, a salt or an ester, and R1 is aC₁-C₅ alkyl group and R2 is a C₂-C₆ alkyl group, may be advantageouslyused for the preparation of a composition intended to modulate themetabolism of lipids in superficial mammalian tissues.

Particularly preferred NMIFAs are those in which R1 is a C₃ alkyl groupand R2 is a C₂-C₆ alkyl group, or in which R2 is a C₄ alkyl group and R1is a C₁-C₅ alkyl group. The most preferred is that in which R1 is ann-propyl group and R2 is an n-butyl group (5,11,14-eicosatrienoic acid,also called 20:3(5,11,14)). The NMIFAs may be preferably provided astriglycerides, phospholipids, fatty acids ester, free fatty acids orcombinations thereof.

Sciadonic acid (5,11,14-eicosatrienoic acid, 20:3Δ5,11,14) is apolyunsaturated fatty acid containing non-methylene-interrupted doublebonds, such as a Δ5-ethylenic bond. Sciadonic acid is often found ingymnosperms, in seed oils, leaves, and wood. It is also found in a fewangiosperms, especially in seed oils. Sciadonic acid has severalbiological activities, including lowering triglyceride and cholesterollevels, reducing reperfusion injury, modifying autoimmune response,having cannabimimetic effect, treatment of skin disease, and treatmentof sensitive or dry skin. WO 95/17987 (The Regents of the University ofCalifornia) shows that broad class of NMIFAs, including5,11,14-eicosatrienoic acid, may be used in an effective amount forsuppressing autoimmune diseases in general, for example rheumatoidarthritis, lupus erythmatosis, multiple sclerosis, myasthenia gravis,and about 30 other diseases currently known. NMIFAs, including5,11,14-eicosatrienoic acid, are further described in U.S. Pat. Nos.5,456,912 and 6,280,755 as well as US Publ. No. 20120156171, each ofwhich is incorporated herein by reference in its entirety.

Pinolenic acid ((5Z,9Z,12Z)-octadeca-5,9,12-trienoic acid;all-cis-5,9,12-18:3) is a fatty acid contained in Siberian Pine nuts,Korean Pine nuts and the seeds of other pines (Pinus species). Thehighest percentage of pinolenic acid is found in Siberian pine nuts andthe oil produced from them.

JP 61 058 536 (Nippon Oil) discloses a method for purifying pine nut oilcontaining at least 10% by weight of 5,9,12-cis-octadecatrienoic acidwhich exhibits a curative effect against arterial hypertension.

WO 96 05 164 (Broadben Nominees Pty) discloses an anti-inflammatorypreparation comprising a purified active fraction, for example5,11,14,17-eicosatetraenoic acid, isolated from a lipid extract of Pernacanalicullus or Mytilus edulis.

Some of the NMIFAs of the invention are naturally occurring substances.Others may be synthesized according to well-known published methodology(see for example Evans et al., Chem. Phys. Lipids, 38, 327-342, 1995).

For example, 20:3(5,11,14) is a naturally occurring substance whichgenerally occurs as one fatty acid in a mixture of fatty acids. ThisNMIFA is found in a wide variety of plants as minor or major fraction ofthe total fatty acid composition. Both the extraction of the mixture offatty acid from their natural sources and the extraction of the20:3(5,11,14) from the resulting fatty acids can be achieved byconventional extraction and purification methods well known among thoseskilled in the art.

The natural sources of fatty acids containing 20:3(5,11,14) areprimarily plant seeds, and prominent among these are conifers andornamental shrubs. The seed oils from these plants are similar to normaledible oils, containing largely oleic, linoleic and linolenic acids, butalso containing useful amounts of NMIFAs. Table 1 lists examples ofseeds whose lipid contents contain significant amounts of 20:3(5,11,14).

% of % of 20:3 (5,11,14) 20:3 (5,11,14) among total among total Sourcefatty acids Source fatty acids Juriperis 14.8 Sciadopitya 15virginiensis verticallara Plarycladus 3 Caltha 23 orientalis palustrisJuriperis 12.3 Calirus 14 chinests rhombuldea Torreya 7 Mortferella 7nucifera alpina* Podocarpus 24 Ephedra 22 nagi campylopoda Anemone 10Anemone 6 rivularis leveilet Cimodefuge 6 Erantis 6 racemosil hyemalisGungko 2.2 Finus 7 biloba silvestria *see the Japanese patent JP5276964(Soatory LTD)

Purification of 20:3(5,11,14) may be in particular achieved by (1)choosing a starting seed source high in total fat content and20:3(5,11,14) content but not containing other contaminating trienes, inparticular alpha-linolenic acid (18:3n-3) and gamma-linolenic acid(18:3n-6) (Podocarpus nagi, Table 1, is such an example); (2) extractingthe lipids with isopropanol and chloroform according to the method ofNichols (Biochim. Biophys Acta 70: 417, 1963); (3) conventionaldegumming and decoloring methods; (4) preparing methyl esters with 2%methanolic sulfuric acid according to the method of Christie (p. 52-53,in Lipid Analysis, Pergamon Press, Oxford, 1982); (5) eluting20:3(5,11,14) methyl ester from a silver nitrate impregnated acid-washedFlorisil column with a hexane:ether mixture ranging from 9:1 to 8:2(volume/volume) according to Carroll, J. Am. Oil Chem. Soc. 40: 413,1963; Wilner, Chem. Ind (Lond) October, 30: 1839, 1965; Merck ChromNews4(1): 1995; Anderson, J. Lipid Res. 6: 577, 1965; and Teshima, Bull.Jap. Soc. Scien. Fish. 44: 927, 1978); (6) removing contaminating silverions by the method of Akesson (Eur. J. Biochem. 9:463, 1969); and (7)optionally converting the methyl ester back to the free acid form bysaponification in 1 M potassium hydroxide in 95% ethanol according toChristie (p. 51-52, in Lipid Analysis, Pergamon Press, Oxford, 1982).

Omega-3 Fatty Acids

Omega 3 fatty acids (also called ω-3 fatty acids or n-3 fatty acids arefats commonly found in marine and plant oils. They are polyunsaturatedfatty acids with a double bond (C═C) starting after the third carbonatom from the end of the carbon chain. The fatty acids have two ends—theacid (COOH) end and the methyl (CH3) end. The location of the firstdouble bond is counted from the methyl end, which is also known as theomega (ω) end or the n end.

Examples of N-3 fatty acids that are important in human physiology areα-linolenic acid (18:3, n-3; ALA), eicosapentaenoic acid (20:5, n-3;EPA), and docosahexaenoic acid (22:6, n-3; DHA). These threepolyunsaturates have either 3, 5, or 6 double bonds in a carbon chain of18, 20, or 22 carbon atoms, respectively. As with mostnaturally-produced fatty acids, all double bonds are in thecis-configuration, in other words, the two hydrogen atoms are on thesame side of the double bond; and the double bonds are methyleneinterrupted, i.e., there are two single bonds between each pair ofadjacent double bonds. Other N-3 fatty acids useful in the presentinvention include:

Hexadecatrienoic acid (HTA); 16:3 (n-3);all-cis-7,10,13-hexadecatrienoic acid

α-Linolenic acid (ALA); 18:3 (n-3); all-cis-9,12,15-octadecatrienoicacid

Stearidonic acid (SDA); 18:4 (n-3); all-cis-6,9,12,15-octadecatetraenoicacid

Eicosatrienoic acid (ETE); 20:3 (n-3); all-cis-11,14,17-eicosatrienoicacid

Eicosatetraenoic acid (ETA); 20:4 (n-3);all-cis-8,11,14,17-eicosatetraenoic acid

Eicosapentaenoic acid (EPA); 20:5 (n-3);all-cis-5,8,11,14,17-eicosapentaenoic acid

Heneicosapentaenoic acid (HPA); 21:5 (n-3;all-cis-6,9,12,15,18-heneicosapentaenoic acid

Docosapentaenoic acid (DPA); 22:5 (n-3);all-cis-7,10,13,16,19-docosapentaenoic acid

Docosahexaenoic acid (DHA); 22:6 (n-3);all-cis-4,7,10,13,16,19-docosahexaenoic acid

Tetracosapentaenoic acid; 24:5 (n-3);all-cis-9,12,15,18,21-tetracosapentaenoic acid

Tetracosahexaenoic acid (Nisinic acid); 24:6 (n-3);all-cis-6,9,12,15,18,21-tetracosahexaenoic acid

In preferred embodiments, the omega-3 fatty acids are marine omega-3fatty acids such as EPA, DHA or DPA. Sources of these fatty acidsinclude, but are not limited to, fish oils (herring oil, salmon oil,tuna oil, anchovy oil, mackerel oil, cod liver oil, sardine oil, and thelike), krill oil, Calanus oil, seal oil, algal oils, bacterial oils,green lipped mussels oil, and the like. Plants may also be geneticallymodified to produce marine omega-3 fatty acids. The marine omega-3 fattyacids may be preferably provided as triglycerides, phospholipids orfatty acids esters or combinations thereof. Herring oil and krill oilare especially preferred sources of marine oil phospholipids. Thetechnology for producing omega-3 concentrates via esterification ofmarine fatty acids and up-concentration by distillation is well known inthe art.

Conjugated Linoleic Acid

In some embodiments, the lipid compositions of the present inventioncomprise one or more conjugated linoleic acid moieties. The conjugatedlinoleic acid moieties may be preferably provided as free fatty acids,esters, acylglycerides or phospholipids. Preferably conjugated linoleicacid isomers include, but are not limited to c9,t11 CLA, t10,c12 CLA,t9,t11 CLA and t10,t12 CLA and combinations thereof. Methods for makingCLA are described in detail in U.S. Pat. Nos. 8,207,225 7,966,0567,776,353 7,514,096 7,452,548 7,115,759 7,094,420 7,078,051 7,029,6916,891,054 6,677,470 6,610,868 6,524,527 6,410,761 6,380,409 6,333,3536,225,486, each or which is incorporated by reference herein in itsentirety.

Non-β-Oxidizable Analogues

The compounds according to the technology may also (especially incombination with sciadonic acid) comprise non-β-oxidizable fatty acidanalogues as represented by the formulaR″CCO—(CH₂)_(2n+1)—X—R′wherein X is a sulfur atom, a selenium atom, an oxygen atom, a CH₂group, a SO group, or a SO₂ group; n is an integer of 0 to 11; R′ is alinear or branched alkyl group, saturated or unsaturated, optionallysubstituted, wherein the main chain of the R′ contains from 13 to 23carbon atoms and optionally one or more heterogroups selected from thegroup comprising an oxygen atom, a sulfur atom, a selenium atom, anoxygen atom, a CH₂ group, a SO group, and a SO₂ group; and R″ is ahydrogen atom or an alkyl group containing from 1 to 4 carbon atoms. Insome preferred embodiments, the non-β-oxidizable fatty acid analogue istetradecylthioacetic acid (TTA), having the structure:

It is understood that analogues that contain one of Se, SO, SO₂, O, orCH₂ in place of sulfur also provide useful pharmaceutical activity. Inaddition, the length and degrees of saturation of the alkyl chains canalso be varied.

The sulfur atom is more electronegative than carbon. Hence, the 3-thiaacid is slightly more acidic than its corresponding fatty acid. Thiafatty acids are also more polar and slightly more soluble in water thanfatty acids of corresponding chain length. Synthetic routes to TTA andmolecules comprising TTA are provided in, e.g., U.S. Pat. Nos.5,093,365; 6,046,237; 6,365,628; 6,417,232; 6,441,036; 7,026,356;7,378,443; 7,902,399; 8,088,825; preparation of mono-, di-, andtriglycerides and nitrogen comprising lipids according to the technologyare disclosed in detail in U.S. Pat. No. 7,375,135; and the synthesis ofphospholipids comprising TTA are provided in, e.g., U.S. Pat. No.8,178,713.

Other Natural Lipids and/or Fatty Acids

While the structure and biological function of many of the major lipidsfrom marine organisms have been studied, less is known of the propertiesof the structure and function of lipids present in marine organisms insmall amounts. Microorganisms such as marine bacteria and algae are theprimary source for lipids and/or lipid precursors in marine food chains.Many potentially bioactive compounds are detectable in fish and othermarine animals in low amounts, e.g., due to losses in the food chain.These bioactive lipids may be present in a larger amount inmicroorganisms nearer the base of the food chain That is, it iscontemplated that lipid extracts (e.g., oils, fractions, etc.)demonstrate high specific biological activities as isolated frommicroorganisms.

For example, furan fatty acids have biological activities such asscavenging free radicals (e.g., by reacting readily with peroxylradicals to generate dioxoenes) and thus may contribute to theprotective properties of fish and fish oil diets relative to heartdisease. Furan fatty acids are tri- or tetra-substituted furanderivatives comprising either a C₃ or C₅ side chain in one of the alphapositions and a straight long-chain saturated acid with a carboxylate atits end in the other alpha position.

Furan fatty acids have been found in fish, algae, bacteria, and fungi,and are generated in large amounts by algae and in small to moderateamounts by plants and other microorganisms. Marine organisms (such asfish) and mammals obtain furan fatty acids in food and metabolize theminto phospholipids. Furan fatty acids are catabolized to dibasicurofuran acids and excreted in the urine. Due to their molecularstructure, furan fatty acids are contemplated to be catabolized moreslowly than other lipids in mammals, and thus potentially to bebioactive in relation to energy metabolism.

As discussed elsewhere, unusual lipids (e.g., phytanyl ether lipids) arepresent in the Archaea, particularly in thermophiles andhyperthermophiles, some of which grow optimally at temperatures higherthan 80° C. In addition, unusual lipids have been found in somethermophilic members of the Bacteria. For example, the lipids found inThermotoga spp. (e.g., T. maritima) comprise a mixture of ether lipidsand ester lipids, mainly polar. Liposomes produced from these lipidsdemonstrate high stability at high and low temperatures, are resistantto acids and bases, and are resistant to high pressure. In addition,these lipids affect membrane properties and thus may result in alteredabsorption and permeability of nutrients.

Bioactive Lipid Compositions

The present invention provides bioactive lipid compositions comprisingone or more bioactive fatty acid moieties, and in particularly preferredembodiments NMIFAs, alone or in combination with other bioactive fattyacid moieties. The bioactive lipid compositions are preferablecharacterized by comprising a particular weight/weight (w/w) percentageof the bioactive fatty acids which refers to the weight of the specificfatty acid or fatty acid moiety as a percentage of the total weight ofthe composition. The bioactive lipid compositions of the presentinvention may comprise free fatty acids, fatty acid esters,monoglycerides, diglycerides, triglycerides, phospholipids andcombinations thereof. Where the bioactive fatty acid is attached to analkyl group, glyceride molecule, or phosphoglyceride molecule via anester or ether bond the fatty acid portion of the molecule is referredto as a fatty acid moiety and the weight percentage of the fatty acidmoiety in the composition is expressed as the weight of the particularfatty acid moiety as a percentage of the total weight of thecomposition.

Thus, the compositions according to the present technology are eitherfatty acids analogous to naturally occurring fatty acids, especiallyNMIFAs alone in combination with other bioactive fatty acids, ornaturally occurring lipids comprising said fatty acid analogues. Invivo, the fatty acid analogues show a strong preference for beingincorporated into phospholipids. Incorporating fatty acid analogues innaturally occurring lipids (e.g., monoglycerides, diglycerides,triglycerides, and/or phospholipids) produces a compound with differentabsorption characteristics compared to the fatty acids. In addition, itis contemplated that incorporating fatty acid analogues in naturallyoccurring lipids (e.g., monoglycerides, diglycerides, triglycerides,and/or phospholipids) may also increase the bioavailability orstability.

For example, some embodiments of the technology relate to atriacylglycerol that includes a NMIFA alone in combination with otherbioactive fatty acids. If such a triacylglycerol were taken orally, forinstance in an animal food product, it would probably be transportedlike any triacylglycerol, e.g., from the small intestine in chylomicronsto the liver; then to the blood in lipoproteins to be stored in theadipose tissue or used by muscles, heart, or the liver; then byhydrolysis of the triacylglycerol into glycerol and three free fattyacids. The free fatty acids would at this point be the fatty acidanalogue parent compound.

Embodiments also encompass glycerophospholipid derivatives of the NMIFAfatty acids, including, but not limited to, phosphatidylcholines,phosphatidylethanolamines, phosphatidylinositols, phosphatidylserines,and phosphatidylglycerols.

In some embodiments, the NMIFAs are incorporated into a sphingolipidderivative such as ceramide or a sphingomyelin. Likeglycerophospholipids complexes, these compounds would be water insolubleand hydrophobic, and thus pass through biological membranes.

Additional embodiments include polar complexes such as, but not limitedto, lysophospholipids, phosphatidic acids, alkoxy compounds,glycerocarbohydrates, gangliosides, and cerebrosides.

Accordingly, in some embodiments, the present invention providesbioactive lipid compositions comprising: a first lipid componentcomprising at least one non-methylene-interrupted fatty acid moiety anda second lipid component comprising at least one bioactive fatty acidmoiety selected from the group consisting of an omega-3 fatty acidmoiety, a non-beta-oxidizable fatty acid moiety, and a conjugatedlinoleic acid moiety. The first and second lipid components maypreferably comprise free fatty acids, acylglycerides, phospholipids,esters and combinations thereof comprising the respective moiety. Insome embodiments, the bioactive composition comprises at least 1% ofsaid first lipid component w/w and at least 1% of said second lipidcomponent w/w. In some embodiments, the composition comprises at least1%, 2%, 3%, 4%, 5%, 10%, 20%, 30%, 40%, 50%, 60%, 70%, 80%, 90%, 95%, or99% w/w of the first lipid component and at least 1%, 2%, 3%, 4%, 5%,10%, 20%, 30%, 40%, 50%, 60%, 70%, 80%, 90%, 95%, or 99% w/w of thesecond lipid component such that the first and second lipid componentsdo not exceed 100% of the composition. In some embodiments, thecompositions comprise at least a third lipid component, an additionalactive component, or a carrier such as a pharmaceutically acceptablecarrier. In these instances the compositions preferably comprise atleast 1%, 2%, 3%, 4%, 5%, 10%, 20%, 30%, 40%, 50%, 60%, 70%, 80%, 90%,95%, or 99% w/w of the first lipid component and at least 1%, 2%, 3%,4%, 5%, 10%, 20%, 30%, 40%, 50%, 60%, 70%, 80%, 90%, 95%, or 99% w/w ofthe second lipid component such that the total amounts of the first andsecond lipid components and at least a third lipid component, additionalactive component and/or carrier do not exceed 100% of the composition.In some embodiments, the compositions comprise a weight ratio ofnon-methylene-interrupted fatty acid:one or more other bioactive fattyacids of from 1:50 to 1:1, from 1:20 to 1:1, from 1:10 to 1:1, from 1:5to 1:1, from 1:4 to 1:1, from 1:3 to 1:1, from 1:2 to 1:1, or from 1.5:1to 1:1. In some embodiments, the compositions comprise a weight ratio ofnon-methylene-interrupted fatty acid:one or more other bioactive fattyacids of from 1:50 to 1:2, from 1:20 to 1:2, from 1:10 to 1:2, from 1:5to 1:2, from 1:4 to 1:2, or from 1:3 to 1:2. In some embodiments, thecompositions comprise a weight ratio of non-methylene-interrupted fattyacid:one or more other bioactive fatty acids of from 1:50 to 1:3, from1:20 to 1:3, from 1:10 to 1:3, from 1:5 to 1:3, or from 1:4 to 1:3. Insome embodiments, the compositions comprise a weight ratio ofnon-methylene-interrupted fatty acid:one or more other bioactive fattyacids of from 1:50 to 1:5, from 1:20 to 1:5, or from 1:10 to 1:5. Insome embodiments, the compositions comprise a weight ratio ofnon-methylene-interrupted fatty acid:one or more other bioactive fattyacids of from 1:50 to 1:10, from 1:40 to 1:10, from 1:30 to 1:10, orfrom 1:20 to 1:10. In some embodiments, the compositions comprise aweight ratio of one or more bioactive fatty acids that are not anon-methylene-interrupted fatty acid:non-methylene-interrupted fattyacid of from 1:50 to 1:2, from 1:20 to 1:2, from 1:10 to 1:2, from 1:5to 1:2, from 1:4 to 1:2, or from 1:3 to 1:2. In some embodiments, thecompositions comprise a weight ratio of one or more bioactive fattyacids that are not a non-methylene-interrupted fattyacid:non-methylene-interrupted fatty acid of from 1:50 to 1:3, from 1:20to 1:3, from 1:10 to 1:3, from 1:5 to 1:3, or from 1:4 to 1:3. In someembodiments, the compositions comprise a weight ratio of one or morebioactive fatty acids that are not a non-methylene-interrupted fattyacid:non-methylene-interrupted fatty acid of from 1:50 to 1:5, from 1:20to 1:5, or from 1:10 to 1:5. In some embodiments, the compositionscomprise a weight ratio of one or more bioactive fatty acids that arenot a non-methylene-interrupted fatty acid:non-methylene-interruptedfatty acid of from 1:50 to 1:10, from 1:40 to 1:10, from 1:30 to 1:10,or from 1:20 to 1:10. In some embodiments, the one or more otherbioactive fatty acid is one or more omega-3 fatty acids. In someembodiments, the omega-3 fatty acid is docosahexaenoic acid (DHA). Insome embodiments, the omega-3 fatty acid is eicosapentaenoic acid (EPA).In some embodiments, the omega-3 fatty acid is docosapentaenoic acid(DPA). In some embodiments, the one or more omega-3 fatty acids are acombination of DHA and EPA. In some embodiments, the one or morebioactive fatty acids is conjugated linoleic acid. In some embodiments,the one or more bioactive fatty acids are a combination of CLA and oneor more omega-3 fatty acids, preferably DHA and EPA. In someembodiments, the one or more bioactive fatty acids is anon-beta-oxidizable fatty acid, preferably tetradecylthioacetic acid(TTA). In some embodiments, the one or more bioactive fatty acids are acombination of a non-beta-oxidizable fatty acid, preferably TTA, and oneor more omega-3 fatty acids, preferably DHA and EPA.

In some embodiments, the non-methylene-interrupted fatty acid moiety isselected from the group consisting of a 5,11,14-eicosatrienoic acidmoiety, a 5,9,12-cis-octadecatrienoic acid moiety; and a5,11,14,17-eicosatetraenoic acid moiety and combinations thereof. Insome embodiments, the omega-3 fatty acid moiety is selected from thegroup consisting of an all-cis-5,8,11,14,17-eicosapentaenoic acidmoiety, an all-cis-7,10,13,16,19-docosapentaenoic acid moiety, and anall-cis-4,7,10,13,16,19-docosahexaenoic acid moiety and combinationsthereof. In some embodiments, the non-beta-oxidizable fatty acid moietyis selected from the group consisting of a tetradecylthioacetic acid(TTA) moiety and a tetradecylselenoacetic acid (TSA) moiety andcombinations thereof. In some embodiments, the conjugated linoleic acidmoiety is selected from the group consisting of a c9,t11 conjugatedlinoleic acid moiety, a t10,c12 conjugated linoleic acid moiety, at9,t11 conjugated linoleic acid moiety, a t10,t12 conjugated linoleicacid moiety and combinations thereof.

The bioactive lipid composition may be preferably provided as an oil,powder, crystal, wax, emulsion, micelle, vesicle, or film. The bioactivelipid composition may be preferably provided in an oral deliveryvehicle, food product, nutritional supplement, dietary supplement orfunctional food.

In some embodiments, the present invention provides structuredphospholipid compositions comprising phospholipid molecules of thefollowing structure:

wherein R1 and R2 are fatty acid moieties or —H and R3 is —H or aphospholipid head group moiety such that the composition comprises atleast 1% w/w of at least one non-methylene-interrupted fatty acid moietyand at least 1% w/w of a second bioactive lipid moiety. In someembodiments, the non-methylene-interrupted fatty acid moiety is selectedfrom the group consisting of a 5,11,14-eicosatrienoic acid moiety, a5,9,12-cis-octadecatrienoic acid moiety; and a5,11,14,17-eicosatetraenoic acid moiety and combinations thereof. Insome embodiments, the bioactive lipid moiety is selected from the groupconsisting of an omega-3 fatty acid moiety, a non-beta-oxidizable fattyacid moiety, a conjugated linoleic acid moiety and combinations thereof.In some embodiments, the omega-3 fatty acid moiety is selected from thegroup consisting of an all-cis-5,8,11,14,17-eicosapentaenoic acidmoiety, an all-cis-7,10,13,16,19-docosapentaenoic acid moiety, and anall-cis-4,7,10,13,16,19-docosahexaenoic acid moiety and combinationsthereof. In some embodiments, the non-beta-oxidizable fatty acid moietyis selected from the group consisting of a tetradecylthioacetic acid(TTA) moiety and a tetradecylselenoacetic acid (TSA) moiety andcombinations thereof. In some embodiments, the conjugated linoleic acidmoiety is selected from the group consisting of a c9,t11 conjugatedlinoleic acid moiety, a t10,c12 conjugated linoleic acid moiety, at9,t11 conjugated linoleic acid moiety, a t10,t12 conjugated linoleicacid moiety and combinations thereof. In some embodiments, thecomposition comprises at least 1%, 2%, 3%, 4%, 5%, 10%, 20%, 30%, 40%,50%, 60%, 70%, 80%, 90%, 95%, or 99% w/w of the at least onenon-methylene-interrupted fatty acid moiety and at least 1%, 2%, 3%, 4%,5%, 10%, 20%, 30%, 40%, 50%, 60%, 70%, 80%, 90%, 95%, or 99% w/w of thesecond bioactive lipid moiety such that the total amounts of therespective non-methylene-interrupted fatty acid moiety and the secondbioactive lipid moiety do not exceed 100% w/w of the composition. Insome embodiments, the compositions comprise at least a third lipidcomponent, an additional active component, or a carrier such as apharmaceutically acceptable carrier. In some embodiments, thephospholipids comprise a weight ratio of non-methylene-interrupted fattyacid:one or more other bioactive fatty acids of from 1:50 to 1:1, from1:20 to 1:1, from 1:10 to 1:1, from 1:5 to 1:1, from 1:4 to 1:1, from1:3 to 1:1, from 1:2 to 1:1, or from 1.5:1 to 1:1. In some embodiments,the compositions comprise a weight ratio of non-methylene-interruptedfatty acid:one or more other bioactive fatty acids of from 1:50 to 1:2,from 1:20 to 1:2, from 1:10 to 1:2, from 1:5 to 1:2, from 1:4 to 1:2, orfrom 1:3 to 1:2. In some embodiments, the compositions comprise a weightratio of non-methylene-interrupted fatty acid:one or more otherbioactive fatty acids of from 1:50 to 1:3, from 1:20 to 1:3, from 1:10to 1:3, from 1:5 to 1:3, or from 1:4 to 1:3. In some embodiments, thecompositions comprise a weight ratio of non-methylene-interrupted fattyacid:one or more other bioactive fatty acids of from 1:50 to 1:5, from1:20 to 1:5, or from 1:10 to 1:5. In some embodiments, the compositionscomprise a weight ratio of non-methylene-interrupted fatty acid:one ormore other bioactive fatty acids of from 1:50 to 1:10, from 1:40 to1:10, from 1:30 to 1:10, or from 1:20 to 1:10. In some embodiments, thecompositions comprise a weight ratio of one or more bioactive fattyacids that are not a non-methylene-interrupted fattyacid:non-methylene-interrupted fatty acid of from 1:50 to 1:2, from 1:20to 1:2, from 1:10 to 1:2, from 1:5 to 1:2, from 1:4 to 1:2, or from 1:3to 1:2. In some embodiments, the compositions comprise a weight ratio ofone or more bioactive fatty acids that are not anon-methylene-interrupted fatty acid:non-methylene-interrupted fattyacid of from 1:50 to 1:3, from 1:20 to 1:3, from 1:10 to 1:3, from 1:5to 1:3, or from 1:4 to 1:3. In some embodiments, the compositionscomprise a weight ratio of one or more bioactive fatty acids that arenot a non-methylene-interrupted fatty acid:non-methylene-interruptedfatty acid of from 1:50 to 1:5, from 1:20 to 1:5, or from 1:10 to 1:5.In some embodiments, the compositions comprise a weight ratio of one ormore bioactive fatty acids that are not a non-methylene-interruptedfatty acid:non-methylene-interrupted fatty acid of from 1:50 to 1:10,from 1:40 to 1:10, from 1:30 to 1:10, or from 1:20 to 1:10. In someembodiments, the one or more other bioactive fatty acid is one or moreomega-3 fatty acids. In some embodiments, the omega-3 fatty acid isdocosahexaenoic acid (DHA). In some embodiments, the omega-3 fatty acidis eicosapentaenoic acid (EPA). In some embodiments, the omega-3 fattyacid is docosapentaenoic acid (DPA). In some embodiments, the one ormore omega-3 fatty acids are a combination of DHA and EPA. In someembodiments, the one or more bioactive fatty acids is conjugatedlinoleic acid. In some embodiments, the one or more bioactive fattyacids are a combination of CLA and one or more omega-3 fatty acids,preferably DHA and EPA. In some embodiments, the one or more bioactivefatty acids is a non-beta-oxidizable fatty acid, preferablytetradecylthioacetic acid (TTA). In some embodiments, the one or morebioactive fatty acids are a combination of a non-beta-oxidizable fattyacid, preferably TTA, and one or more omega-3 fatty acids, preferablyDHA and EPA.

Accordingly, articular embodiments relate to phospholipids comprisingone or more of a non-methylene-interrupted fatty acid moiety and asecond bioactive lipid moiety, for example, an omega-3 fatty acidmoiety, a non-beta-oxidizable fatty acid moiety, and a conjugatedlinoleic acid moiety. Phospholipids according to the technology are notlimited in the polar headgroup of the phospholipid. For example, thepolar head group may be the polar head group of any suitable lipid.

In some embodiments, the phospholipid is a neutral or anionicphospholipid. For example, in some embodiments the polar head group isthe polar head group of, or is derived from, a lipid such as aphospholipid, ceramide, triacylglycerol, lysophospholipid,phosphatidylserine, glycerol, alcohol, alkoxy compound,monoacylglycerol, ganglioside, sphingomyelin, cerebroside,phosphatidylcholine (e.g., dioleoylphosphatidylcholine (DOPC)),phosphatidylethanolamine (e.g., dioleoylphosphatidylethanolamine(DOPE)), phosphatidylinositol, diacylglycerol, phosphatidic acid,glycerocarbohydrate, polyalcohol, and/or phosphatidylglycerol.

Exemplary polar headgroups are, e.g.:

In some embodiments, the polar head group is, or is derived from, atriacylglycerol, e.g., having the structure

In some embodiments, the polar head group (PHG) comprises the group—W-Linker-HGwherein W is selected from CH₂, O, NR¹, and S, wherein R¹ is H or ahydrocarbyl group, wherein Linker is an optional linker group, and HG isa head group.

The head group (HG) may be polar or non-polar. When HG is non-polar itmay be rendered polar by group —C(O)W-Linker-. Such head groups areencompassed by the present definition provided —C(O)W-Linker-HG is polarand HG is polar when attached to the —C(O)W-Linker-group.

In some embodiments, the head group (HG) may be an alkyl group, e.g.,having at least 5 carbons. In some embodiments, it is a C₅₋₁₀₀ alkylgroup, a C₅₋₈₀ alkyl group, a C₅₋₆₀ alkyl group, a C₅₋₅₀ alkyl group, aC₅₋₄₀ alkyl group, C₅₋₃₀ alkyl group, or a C₅₋₂₀ alkyl group.

For example, in some embodiments the HG is the head group of, or isderived from, a lipid such as a phospholipid, ceramide, triacylglycerol,lysophospholipid, phosphatidylserine, glycerol, alcohol, alkoxycompound, monoacylglycerol, ganglioside, sphingomyelin, cerebroside,phosphatidylcholine, phosphatidylethanolamine, phosphatidylinositol,diacylglycerol, phosphatidic acid, glycerocarbohydrate, polyalcohol,and/or a phosphatidylglycerol.

Exemplary head groups are, e.g.:

The structured phospholipid composition may be preferably provided as anoil, powder, crystal, wax, emulsion, micelle, vesicle, or film. Thestructured phospholipid composition may be preferably provided in anoral delivery vehicle, food product, nutritional supplement, dietarysupplement or functional food.

In some embodiments, the present invention provides a structuredacylglycerol composition comprising acylglycerol molecules of thefollowing structure:

wherein R1, R2 and R3 are fatty acid moieties or —H such that saidcomposition comprises at least 1% w/w of at least onenon-methylene-interrupted fatty acid moiety and at least 1% w/w of asecond bioactive lipid moiety. In some embodiments, thenon-methylene-interrupted fatty acid moiety is selected from the groupconsisting of a 5,11,14-eicosatrienoic acid moiety, a5,9,12-cis-octadecatrienoic acid moiety; and a5,11,14,17-eicosatetraenoic acid moiety and combinations thereof. Insome embodiments, the bioactive lipid moiety is selected from the groupconsisting of an omega-3 fatty acid moiety, a non-beta-oxidizable fattyacid moiety, a conjugated linoleic acid moiety and combinations thereof.In some embodiments, the omega-3 fatty acid moiety is selected from thegroup consisting of an all-cis-5,8,11,14,17-eicosapentaenoic acidmoiety, an all-cis-7,10,13,16,19-docosapentaenoic acid moiety, and anall-cis-4,7,10,13,16,19-docosahexaenoic acid moiety and combinationsthereof. In some embodiments, the non-beta-oxidizable fatty acid moietyis selected from the group consisting of a tetradecylthioacetic acid(TTA) moiety and a tetradecylselenoacetic acid (TSA) moiety andcombinations thereof. In some embodiments, the conjugated linoleic acidmoiety is selected from the group consisting of a c9,t11 conjugatedlinoleic acid moiety, a t10,c12 conjugated linoleic acid moiety, at9,t11 conjugated linoleic acid moiety, a t10,t12 conjugated linoleicacid moiety and combinations thereof. In some embodiments, thecomposition comprises at least 1%, 2%, 3%, 4%, 5%, 10%, 20%, 30%, 40%,50%, 60%, 70%, 80%, 90%, 95%, or 99% w/w of the at least onenon-methylene-interrupted fatty acid moiety and at least 1%, 2%, 3%, 4%,5%, 10%, 20%, 30%, 40%, 50%, 60%, 70%, 80%, 90%, 95%, or 99% of thesecond bioactive lipid moiety such that the total amounts of therespective non-methylene-interrupted fatty acid moiety and the secondbioactive lipid moiety do not exceed 100% w/w of the composition. Insome embodiments, the compositions comprise at least a third lipidcomponent, an additional active component, or a carrier such as apharmaceutically acceptable carrier. In some embodiments, theacylglycerols, preferably triglycerides, comprise a weight ratio ofnon-methylene-interrupted fatty acid:one or more other bioactive fattyacids of from 1:50 to 1:1, from 1:20 to 1:1, from 1:10 to 1:1, from 1:5to 1:1, from 1:4 to 1:1, from 1:3 to 1:1, from 1:2 to 1:1, or from 1.5:1to 1:1. In some embodiments, the compositions comprise a weight ratio ofnon-methylene-interrupted fatty acid:one or more other bioactive fattyacids of from 1:50 to 1:2, from 1:20 to 1:2, from 1:10 to 1:2, from 1:5to 1:2, from 1:4 to 1:2, or from 1:3 to 1:2. In some embodiments, thecompositions comprise a weight ratio of non-methylene-interrupted fattyacid:one or more other bioactive fatty acids of from 1:50 to 1:3, from1:20 to 1:3, from 1:10 to 1:3, from 1:5 to 1:3, or from 1:4 to 1:3. Insome embodiments, the compositions comprise a weight ratio ofnon-methylene-interrupted fatty acid:one or more other bioactive fattyacids of from 1:50 to 1:5, from 1:20 to 1:5, or from 1:10 to 1:5. Insome embodiments, the compositions comprise a weight ratio ofnon-methylene-interrupted fatty acid:one or more other bioactive fattyacids of from 1:50 to 1:10, from 1:40 to 1:10, from 1:30 to 1:10, orfrom 1:20 to 1:10. In some embodiments, the compositions comprise aweight ratio of one or more bioactive fatty acids that are not anon-methylene-interrupted fatty acid:non-methylene-interrupted fattyacid of from 1:50 to 1:2, from 1:20 to 1:2, from 1:10 to 1:2, from 1:5to 1:2, from 1:4 to 1:2, or from 1:3 to 1:2. In some embodiments, thecompositions comprise a weight ratio of one or more bioactive fattyacids that are not a non-methylene-interrupted fattyacid:non-methylene-interrupted fatty acid of from 1:50 to 1:3, from 1:20to 1:3, from 1:10 to 1:3, from 1:5 to 1:3, or from 1:4 to 1:3. In someembodiments, the compositions comprise a weight ratio of one or morebioactive fatty acids that are not a non-methylene-interrupted fattyacid:non-methylene-interrupted fatty acid of from 1:50 to 1:5, from 1:20to 1:5, or from 1:10 to 1:5. In some embodiments, the compositionscomprise a weight ratio of one or more bioactive fatty acids that arenot a non-methylene-interrupted fatty acid:non-methylene-interruptedfatty acid of from 1:50 to 1:10, from 1:40 to 1:10, from 1:30 to 1:10,or from 1:20 to 1:10. In some embodiments, the one or more otherbioactive fatty acid is one or more omega-3 fatty acids. In someembodiments, the omega-3 fatty acid is docosahexaenoic acid (DHA). Insome embodiments, the omega-3 fatty acid is eicosapentaenoic acid (EPA).In some embodiments, the omega-3 fatty acid is docosapentaenoic acid(DPA). In some embodiments, the one or more omega-3 fatty acids are acombination of DHA and EPA. In some embodiments, the one or morebioactive fatty acids is conjugated linoleic acid. In some embodiments,the one or more bioactive fatty acids are a combination of CLA and oneor more omega-3 fatty acids, preferably DHA and EPA. In someembodiments, the one or more bioactive fatty acids is anon-beta-oxidizable fatty acid, preferably tetradecylthioacetic acid(TTA). In some embodiments, the one or more bioactive fatty acids are acombination of a non-beta-oxidizable fatty acid, preferably TTA, and oneor more omega-3 fatty acids, preferably DHA and EPA.

The structured acylglycerol composition may be preferably provided as anoil, powder, crystal, wax, emulsion, micelle, vesicle, or film. Thestructured phospholipid composition may be preferably provided in anoral delivery vehicle, food product, nutritional supplement, dietarysupplement or functional food.

In some embodiments, the present invention provides bioactive lipidcompositions comprising at least 1%, 2%, 3%, 4%, 5%, 10%, 20%, 30%, 40%,50%, 60%, 70%, 80%, 90%, 95%, or 99% w/w of the structured phospholipidcomposition described above and at least at least 1%, 2%, 3%, 4%, 5%,10%, 20%, 30%, 40%, 50%, 60%, 70%, 80%, 90%, 95%, or 99% w/w of thestructured acylglycerol composition described above such that the totalamount of the respective compositions does not exceed 100%.

The present invention likewise provides methods of using thecompositions. These methods and uses are described in detail below butmay be summarized as follows. In some embodiments, the present inventionprovides methods of treating a subject comprising administering to saidsubject the bioactive lipid composition, structured phospholipidcomposition or structured acylglyceride composition or oral deliveryvehicle, food product, nutritional supplement, dietary supplement orfunction food as described above to a subject in need thereof. In someembodiments, the administration or oral, topical, parenteral, enteral,transdermal, intradermal, intraocular, intravitreal, sublingual, orintravaginal and may preferably comprise an effective amount of thecomposition.

In further preferred embodiments, the present invention provides methodsof reducing obesity, inducing weight loss, increasing lean body mass,increasing muscularity, increasing muscle mass, improving bodycomposition, alleviating one or more symptoms metabolic syndrome,treating diabetes, decreasing insulin resistance, reducing inflammation,improving concentration, memory, cognitive function, attention andtreating, alleviating or improving one or more of the following diseasesor conditions: re stenosis, arteriosclerosis, coronary heart disease,thrombosis, myocardial infarction, stroke, hypertension, fatty liver,diabetes, hyperglycaemia, hyperinsulinemia, and stenosis, rheumatoidarthritis, systemic vasculitis, systemic lupus erythematosus, systemicsclerosis, dermatomyositis, polymyositis, various autoimmune endocrinedisorders (e.g. thyroiditis and adrenalitis), various immune mediatedneurological disorders (e.g. multiple sclerosis and myastenia gravis),various cardiovascular disorders (e.g. myocarditis, congestive heartfailure, arteriosclerosis and stable and unstable angina, and Wegenersgranulomatosis), inflammatory bowel diseases and colitis (e.g., Crohn'scolitis), nephritis, various inflammatory skin disorders (e.g.psoriasis, atopic dermatitis and food allergy) and acute and chronicallograft rejection after organ transplantation, comprisingadministering to a subject in need thereof the bioactive lipidcomposition, structured phospholipid composition or structuredacylglyceride composition or oral delivery vehicle, food product,nutritional supplement, dietary supplement or function food as describedabove. In some embodiments, the administration or oral, topical,parenteral, enteral, transdermal, intradermal, intraocular,intravitreal, sublingual, or intravaginal and may preferably comprise aneffective amount of the composition. The treatment is preferablyperformed under conditions such that the disease or condition isalleviated or improved.

Transesterification

In some embodiments, the lipid compositions are made bytransesterification. In some preferred embodiments, a natural startingoil is used. Examples of suitable starting oils include Korean pine oil,Siberian pine oil, and oils from other sources identified in Table 1above, fish oil with a high triglyceride content, a fish oil with a highphospholipid content (e.g., herring oil), a krill oil with a highphospholipid content, and marine oil concentrates comprising esters,fatty acids or triglycerides with contents of EPA and/or DHA.Transesterification is preferably used to replace fatty acids in thestarting oil with a desired fatty acid. For example, a starting oilcomprising a non-methylene-interrupted fatty acid moiety such as a pineoil may be transesterified with a second lipid composition comprising adesired omega-3 fatty acid moiety, a non-beta-oxidizable fatty acidmoiety, or conjugated linoleic acid moiety. As another example, astarting marine oil with a high phospholipid content may betransesterified with a suitable source of non-methylene-interruptedfatty acid moieties. Methods for transesterification are provided inU.S. Publ. No. 20060177486 and 20030144353, each of which isincorporated herein by reference.

In some embodiments, novel acylglycerides of the present invention aremanufactured by using non-specific and position-specific lipases toinsert a first fatty acyl residue at position 2 (SN2) of theacylglyceride and a second fatty acyl residue at positions 1 and 3 (SN1and SN3) of the acylglyceride. Non-specific lipases are lipases that areable to hydrolyse or esterify (i.e., the reverse reaction) fatty acidsin all positions on a glycerol. A position-specific or 1,3 specificlipase almost exclusively hydrolyses or esterifies fatty acids inposition 1 and 3 on the glycerol backbone. The structured acylglyceridesof the present invention are synthesized by first using a non-specificlipase to attach the desired fatty acid for position 2 to all 3positions and then hydrolysing the acyl residues in position 1 and 3using a 1,3 specific lipase. The hydrolysed acids are then removed bydistillation before the acids desired to be attached to positions 1 are3 are added and esterified to position 1 and 3 by the same lipase. Thedirection of the reaction (hydrolysis or esterification) is easilycontrolled by water addition or removal respectively. In the followingexample is a general outline of the method.

In particularly preferred embodiments, a purified aliquot of a firstfatty acid (about 3 moles), glycerol (about 1 mole) and up to 10% byweight of acids are mixed with immobilized non-specific lipase(commercially available). The mixture is stirred under vacuum andslightly heated (50-60 C). The water produced during the esterificationis continuously removed by the vacuum suction. After 24-48 hours, thereaction is finished and the enzymes are removed and recovered byfiltration. The resulting acylglyceride has the first fatty acidattached at all three positions. The first fatty acid residue atpositions 1 and 3 is then removed in by addition of 1,3 specificimmobilized lipase (commercially available) and 1% water. The mixture isheated to 50-60 C. and stirred under nitrogen atmosphere for 24-48hours. The reaction mixture now comprises free fatty acids liberatedfrom position 1 and 3 and monoglycerides (fatty acid B attached toposition 2). Next, in preferred embodiments, the fatty acids aredistilled off from the mixture by molecular distillation. In furtherpreferred embodiments, about one mole of the monoglyceride is allowed toreact for 24-48 hours with 2 moles a second free fatty acid in thepresence of 1,3 specific lipase. In some embodiments, this reactiontakes place under stirring and vacuum at 50-60.degree. C. to removewater produced in the esterification process. The resultingacylglyceride is a structured triglyceride with the first fatty acid inposition 2 and the second fatty acid in positions 1 and 3.

As described above, in some embodiments of the present invention, lipasethat specifically acts on the positions 1 and 3 of triglyceride is usedas catalyst. The present invention is not limited to the use of anyparticular 1,3 specific lipase. Examples of 1,3 specific lipases usefulin the present invention include lipases produced by a microorganismbelonging to the genus Rhizopus, Rhizomucor, Mucor, Penicillium,Aspergillus, Humicola or Fusarium, as well as porcine pancreatic lipase.Examples of commercially available lipases include lipase of Rhizopusdelemar (Tanabe Pharmaceutical, Dalipase), lipase of Rhizomucor miehei(Novo Nordisk, Ribozyme IM), lipase of Aspergillus niger (AmanoPharmaceutical, Lipase A), lipase of Humicola lanuginosa (Novo Nordisk,Lipolase), lipase of Mucor javanicus (Amano Pharmaceutical, Lipase M)and lipase of Fusarium heterosporum. These lipases may be used in theirnative form, or in the form of lipase that has been immobilized oncellite, ion exchange resin or a ceramic carrier.

The amount of water added to the reaction system affects the outcome ofthe reaction. Transesterification does not proceed in the absoluteabsence of water, while if the amount of water is too much, hydrolysisoccurs, the triglyceride recovery rate decreases, or spontaneous acylgroup transfer occurs in a partially acylated glyceride resulting intransfer of the saturated fatty acid at the position 2 to the position 1or 3. Thus, when using an immobilized enzyme that does not have bondedwater, it is effective to first activate the enzyme using a substrate towhich water has been added before carrying out the reaction, and thenuse a substrate to which water is not added during the reaction. Inorder to activate the enzyme in batch reactions, a substrate containingwater at 0 to 1,000% (wt %) of the amount of added enzyme should be usedto pretreat the enzyme, and in the case of activating by a columnmethod, a water-saturated substrate should be allowed to continuouslyflow through the column. The amount of lipase used in a batch reactionmay be determined according to the reaction conditions. Although thereare no particular limitations on the amount of lipase, 1 to 30% (wt %)of the reaction mixture is suitable when using, for example, lipase ofRhizopus delemar or lipase of Rhizomucor miehei immobilized on celliteor a ceramic carrier.

In some preferred embodiments, the above-mentioned immobilized enzymecan be used repeatedly. Namely, the reaction can be continued by leavingthe immobilized enzyme in a reaction vessel after reaction and replacingthe reaction mixture with freshly prepared reaction mixture comprisingsubstrate. In addition, for transesterification by a column method, areaction mixture containing substrate be allowed to flow continuously atthe rate of 0.05 to 20 ml/hr per gram of enzyme. In other preferredembodiments, the content of target triglyceride can be increased byperforming transesterification repeatedly. Namely, lipase specificallyacting on the positions 1 and 3 of the acylglyceride is allowed to actin the presence of the second fatty acid or an ester thereof to obtain areaction mixture in which fatty acids at positions 1 and 3 aretransesterified to the desired fatty acid.

The target acylglycerides of the present invention can easily beisolated by routine methods such as liquid chromatography, moleculardistillation, downstream membrane fractionation or vacuumsuperfractionation or a combination thereof. Purification of the targetacylglycerides of the present invention can be performed by alkalinedeacidation, steam distillation, molecular distillation, downstreammembrane fractionation, vacuum superfractionation, columnchromatography, solvent extraction or membrane separation, or acombination thereof so as to remove the above-mentioned fatty acidsreleased by the transesterification and unreacted unsaturated fattyacids.

In some embodiments, the present invention utilizes a phospholipid,preferably a phosphatide such as lecithin (e.g., egg lecithin, krilllecithin, herring lecithin, soybean lecithin, or egg lecithin), in anenzymatic reaction so that the fatty acid in position 1 of thephospholipid is replaced with a desired fatty acid residue. The presentinvention is not limited to the use of any particular phospholipid.Indeed, the use of a variety of phospholipids is contemplated. In someembodiments, the phospholipid is a phosphatidic or lysophosphatidicacid. In more preferred embodiments, the phospholipid is a mixture ofphosphatides such as phosphatidylcholine, phosphatidylethanolamine,phosphatidylserine and phosphatidylinositol. The present invention isnot limited to the use of any particular source of phospholipids.

In preferred embodiments, the replacement (e.g., by transesterification)of the phospholipid fatty acids with a desired fatty acid or theaddition (e.g. esterification) is catalyzed by a lipase. The presentinvention is not limited to the use of any particular lipase. Indeed,the use of a variety of lipases is contemplated, including, but notlimited to, the aforementioned Thermomyces Lanuginosus lipase,Rhizomucor miehei lipase, Candida Antarctica lipase, Pseudomonasfluorescence lipase, and Mucor javanicus lipase. It is contemplated thata variety of desired fatty acids may be substituted onto thephospholipids utilized in the process of the present invention,especially fatty acids that are not initially present in the startingphospholipid composition such as a non-methylene-interrupted fatty acidmoiety and a second bioactive lipid moiety, an omega-3 fatty acidmoiety, a non-beta-oxidizable fatty acid moiety, a conjugated linoleicacid moiety and combinations thereof.

Chemical Synthesis

Acylation of sn-glycero-3-phosphocholine (GPC) with an activated fattyacid, such as fatty acid imidazolides, is a standard procedure inphosphatidylcholine synthesis. It is usually carried out in the presenceof DMSO anion with DMSO as solvent (Hermetter; Chemistry and Physics oflipids, 1981, 28, 111). Sn-Glycero-3-phosphocholine, as cadmium (II)adduct can also be reacted with the imidazolide activated fatty acid inthe presence of DBU (1,8-diazabicyclo[5.4.0]undec-7-ene] to prepare thephosphatidylcholine of the respective fatty acid (Internationalapplication number PCT/GB2003/002582). Enzymatic transphosphatidylationcan affect the transformation of phosphatidylcholine tophosphatidylethanolamine (Wang et al, J. Am. Chem. Soc., 1993, 115,10487). In other embodiments, a lysophospholipid with a desiredbioactive fatty acid moiety (e.g., omega-3 fatty acid moiety, conjugatedlinoleic acid moiety of sciadonic acid moiety) at the SN-1 or SN-2position is acylated with a non-beta-oxidizable fatty acid analoguemoiety by combining desired omega-3 fatty acid non-beta-oxidizable fattyacid analogue moiety anhydride (e.g. from TTA) and 4-pyrrolidinopyridineas a catalyst (1.2 equivalents) in alcohol-free chloroform.Polyunsaturated fatty acids containing phospholipids may be prepared byvarious ways, mainly by chemical synthesis of phospholipids asdescribed, by enzymatic esterification and transesterification ofphospholipids or enzymatic transphosphatidylation of phospholipids.(Hosokawa, J. Am. Oil Chem. Soc. 1995, 1287, Lilja-Hallberg,Biocatalysis, 1994, 195).

Isolation of Phospholipids from a Living Organism, Consortium, or System

In some embodiments, the technology relates to isolating phospholipids(e.g., a natural phospholipid and/or an oil) comprising a desired fattyacid moiety or combination of fatty acid moieties (e.g.,non-methylene-interrupted fatty acid moiety, an omega-3 fatty acidmoiety, a non-beta-oxidizable fatty acid moiety, a conjugated linoleicacid moiety and combinations thereof) from one or more living organismssuch as a bacterium, alga, archaeon, yeast, etc. Phospholipids make upapproximately 10% of the dry weight of a cell. Thus, cultures of cellsprovide a source for the production and harvesting of phospholipids.Accordingly, in some embodiments, the technology relates to feeding aliving organism a desired fatty acid moiety or combination, which isthen incorporated into phospholipid by the living organism, and thenisolating the phospholipid, e.g., by biochemical or other isolationand/or purification techniques. In some embodiments, the phospholipid isco-isolated with other biological molecules, substances, entities, etc.that are produced by the living organism. That is, in some embodiments,the technology relates to a composition (e.g., an oil) and/or a methodof producing a composition produced from a living organism, wherein thecomposition comprises a phospholipid (e.g., a natural phospholipid)having one or more desired fatty acid moieties(non-methylene-interrupted fatty acid moiety, an omega-3 fatty acidmoiety, a non-beta-oxidizable fatty acid moiety, a conjugated linoleicacid moiety and combinations thereof) and optionally at least one otherbiological molecule from the living organism.

The living organism may, in some embodiments, be grown in a controlledculture, e.g., in a defined medium, a semi-defined medium, an undefinedmedium, a synthetic medium, or a natural medium; under controlledtemperature, pressure, volume, and agitation; and in a controlledatmosphere of gases (e.g., a particular mixture of oxygen, carbondioxide, nitrogen, and other gases, etc.). The culture may comprise asingle type of organism (e.g., a single species, sub-species, clone,subtype, isolate, etc.) or the culture may comprise more than one typeof organism (e.g., more than one species, sub-species, clone, subtype,isolate, etc.).

In some embodiments, the living organism is a member of the Bacteria; insome embodiments, the living organism is a member of the Eukarya; and,in some embodiments, the living organism is a member of the Archaea, asdefined by, e.g., Woese C, Fox G (1977). “Phylogenetic structure of theprokaryotic domain: the primary kingdoms.” Proc Natl Acad Sci USA 74:5088-90; Woese C, Kandler O, Wheelis M (1990). “Towards a natural systemof organisms: proposal for the domains Archaea, Bacteria, and Eucarya.”Proc Natl Acad Sci USA 87: 4576-9. In some embodiments, one or morephospholipids according to the technology is or are isolated from acomposition of more than one living organism, e.g., a co-culture and/orliving system and/or consortium of living organisms that may or may notbe categorized in the same phylogenetic kingdom.

It is contemplated that any organism that can be grown in the presenceof the desired fatty acid moiety is encompassed by the presenttechnology. It is contemplated that any organism that can incorporate adesired fatty acid moiety into a phospholipid is encompassed by thepresent technology.

Further Aspects

In some embodiments, the compositions provided herein are combined witha liposome or formulated into a micellar form to assist inadministration. In some embodiments, compounds are formulated in acochleate delivery vehicle. Cochleate delivery vehicles are a newtechnology platform for oral delivery of drugs. Cochleates are stablephospholipid-cation precipitates composed of simple, naturally occurringmaterials, for example, phosphatidylserine and calcium. Cochleates are apotential nanosized system that can encapsulate hydrophobic,amphiphilic, negatively, or positively charged moieties.

In some embodiments, the compound is an isolated form or purified form.For example, the compound may be in a form or at a purity other thanthat found in a biological system such as in vivo. In some embodiments,the compound is semi-isolated or semi-purified, e.g., the compound is anisolated form or purified form and is present in a composition with oneor more other biological molecules that are not contaminants orimpurities. In some embodiments, the compounds provided are formulatedto provide a pharmaceutical composition comprising a compound accordingto the technology and/or a pharmaceutically acceptable carrier, diluent,excipient, or adjuvant.

An addition, embodiments provide compounds that are lipids comprising atleast one non-polar moiety and a polar moiety, wherein the non-polarmoiety is of the formula X—Y—Z—, wherein X is a hydrocarbyl chain; Y isS, Se, SO₂, SO, O, or CH₂; and Z is an optional hydrocarbyl group.Furthermore, when Y is CH₂, the chain X—Y—Z contains an even number ofatoms, the polar moiety is —[C(O)]_(m)PHG, wherein PHG is a polar headgroup and m is the number of non-polar moieties.

Pharmaceutical Compositions

Provided herein are pharmaceutical compositions comprising atherapeutically effective amount of a composition according to thepresent technology and a pharmaceutically acceptable carrier, diluent,or excipient (including combinations thereof).

A composition according to the technology comprises or consists of atherapeutically effective amount of a pharmaceutically active agent. Insome embodiments, it includes a pharmaceutically acceptable carrier,diluent, or excipient (including combinations thereof). Acceptablecarriers or diluents for therapeutic use are well known in thepharmaceutical art and are described, for example, in Remington'sPharmaceutical Sciences, Mack Publishing Co. (A. R. Gennaro edit. 1985).The choice of pharmaceutical carrier, excipient, or diluent is selectedwith regard to the intended route of administration and standardpharmaceutical practice. The pharmaceutical comprise as, or in additionto, the carrier, excipient, or diluent any suitable binder(s),lubricant(s), suspending agent(s), coating agent(s), and/or solubilizingagent(s).

This pharmaceutical composition will desirably be provided in a sterileform. It may be provided in unit dosage form and will generally beprovided in a sealed container. A plurality of unit dosage forms may beprovided.

Pharmaceutical compositions within the scope of the present technologymay include one or more of the following: preserving agents,solubilizing agents, stabilizing agents, wetting agents, emulsifiers,sweeteners, colorants, flavoring agents, odorants, and/or salts.Compounds of the present technology may themselves be provided in theform of a pharmaceutically acceptable salt. In addition, embodiments maycomprise buffers, coating agents, antioxidants, suspending agents,adjuvants, excipients, and/or diluents. Examples of preservativesinclude sodium benzoate, sorbic acid, and esters of p-hydroxybenzoicacid.

They may also contain other therapeutically active agents in addition tocompounds of the present technology. Where two or more therapeuticagents are used they may be administered separately (e.g., at differenttimes and/or via different routes) and therefore do not always need tobe present in a single composition. Thus, combination therapy is withinthe scope of the present technology.

Route of Administration

A pharmaceutical composition within the scope of the present technologymay be adapted for administration by any appropriate route. For example,it may be administered by the oral (including buccal or sublingual),rectal, nasal, topical (including buccal, sublingual, or transdermal),vaginal, or parenteral (including subcutaneous, intramuscular,intravenous, or intradermal) routes. Such a composition may be preparedby any method known in the art of pharmacy, for example, by admixing oneor more active ingredients with a suitable carrier.

In various embodiments, different drug delivery systems are used toadminister pharmaceutical compositions of the present technology,depending upon the desired route of administration. Drug deliverysystems are described, for example, by Langer (Science 249:1527-1533(1991)) and by Illum and Davis (Current Opinions in Biotechnology 2:254-259 (1991)).

The agents of the present technology may be administered alone but willgenerally be administered as a pharmaceutical composition—e.g., theagent is in admixture with a suitable pharmaceutical excipient, diluent,or carrier selected with regard to the intended route of administrationand standard pharmaceutical practice. For example, in some embodimentsthe agent is administered (e.g., orally or topically) in the form oftablets, capsules, ovules, elixirs, solutions, or suspensions, which maycontain flavoring or coloring agents, for immediate, delayed, modified,sustained, pulsed, and/or controlled-release applications.

In some embodiments, tablets contain excipients such as microcrystallinecellulose, lactose, sodium citrate, calcium carbonate, dibasic calciumphosphate and/orglycine; disintegrants such as starch (preferably corn,potato, or tapioca starch), sodium starch glycollate, croscarmellosesodium, and/or certain complex silicates; and/or granulation binderssuch as polyvinylpyrrolidone, hydroxypropylmethylcellulose (HPMC),hydroxypropylcellulose (HPC), sucrose, gelatin, and/or acacia.Additionally, lubricating agents such as magnesium stearate, stearicacid, glyceryl behenate, and talc may be included.

In some embodiments, solid compositions of a similar type are alsoemployed as fillers in gelatin capsules. Examples of excipients in thisregard include lactose, starch, a cellulose, milk sugar, or highmolecular weight polyethylene glycols. For some embodiments of aqueoussuspensions and/or elixirs, the agent is combined with varioussweetening or flavoring agents, coloring matter or dyes, withemulsifying and/or suspending agents and with diluents such as water,ethanol, propylene glycol, and glycerin, and combinations thereof.

The routes for administration (delivery) include, but are not limitedto, one or more of: oral (e.g. as a tablet, capsule, or as an ingestablesolution), topical, mucosal (e.g. as a nasal spray or aerosol forinhalation), nasal, parenteral (e.g. by an injectable form),gastrointestinal, intraspinal, intraperitoneal, intramuscular,intravenous, intrauterine, intraocular, intradermal, intracranial,intratracheal, intravaginal, intracerebroventricular, intracerebral,subcutaneous, ophthalmic (including intravitreal or intracameral),transdermal, rectal, buccal, via the penis, vaginal, epidural,sublingual.

It is to be understood that not all of the agent need be administered bythe same route. Likewise, if the composition comprises more than oneactive component, then those components may be administered by differentroutes.

If the agent of the present technology is administered parenterally,then examples of such administration include one or more of:intravenously, intra-arterially, intraperitoneally, intrathecally,intraventricularly, intraurethrally, intrastemally, intracranially,intramuscularly, or subcutaneously administering the agent; and/or byusing infusion techniques.

Oral Administration

In some embodiments, pharmaceutical compositions adapted for oraladministration are provided as capsules or tablets; as powders orgranules; as solutions, syrups or suspensions (in aqueous or non-aqueousliquids); as edible foams or whips; or as emulsions. Tablets or hardgelatine capsules may comprise lactose, maize starch or derivativesthereof, stearic acid or salts thereof. Soft gelatine capsules maycomprise vegetable oils, waxes, fats, semi-solid, or liquid polyols etc.Solutions and syrups may comprise water, polyols and sugars. For thepreparation of suspensions, oils (e.g., vegetable oils) may be used toprovide oil-in-water or water-in-oil suspensions. An active agentintended for oral administration may be coated with or admixed with amaterial that delays disintegration and/or absorption of the activeagent in the gastrointestinal tract (e.g., glyceryl monostearate orglyceryl distearate may be used). Thus, the sustained release of anactive agent may be achieved over many hours and, if necessary, theactive agent can be protected from being degraded within the stomach.Pharmaceutical compositions for oral administration may be formulated tofacilitate release of an active agent at a particular gastrointestinallocation due to specific pH or enzymatic conditions.

Transdermal Administration

Pharmaceutical compositions adapted for transdermal administration maybe provided as discrete patches intended to remain in intimate contactwith the epidermis of the recipient for a prolonged period of time. Forexample, the active ingredient may be delivered from the patch byiontophoresis, e.g., as described in Pharmaceutical Research, 3: 318(1986)).

Topical Administration

Alternatively, the agent of the present technology can be administeredin the form of a suppository or pessary, or it may be applied topicallyin the form of a gel, hydrogel, lotion, solution, cream, ointment ordusting powder. The agent of the present technology may also be dermallyor transdermally administered, for example, by the use of a skin patch.They may also be administered by the pulmonary or rectal routes. Theymay also be administered by the ocular route. For ophthalmic use, thecompounds can be formulated as micronised suspensions in isotonic, pHadjusted, sterile saline, or, preferably, as solutions in isotonic, pHadjusted, sterile saline, optionally in combination with a preservativesuch as a benzylalkonium chloride. Alternatively, they may be formulatedin an ointment such as petrolatum.

For application topically to the skin, the agent of the presenttechnology can be formulated as a suitable ointment containing theactive compound suspended or dissolved in, for example, a mixture withone or more of the following: mineral oil, liquid petrolatum, whitepetrolatum, propylene glycol, polyoxyethylene polyoxypropylene compound,emulsifying wax and water. Alternatively, it can be formulated as asuitable lotion or cream, suspended or dissolved in, for example, amixture of one or more of the following: mineral oil, sorbitanmonostearate, a polyethylene glycol, liquid paraffin, polysorbate 60,cetyl esters wax, cetearyl alcohol, 2-octyldodecanol, benzyl alcohol andwater.

Rectal Administration

Pharmaceutical compositions adapted for rectal administration may beprovided as suppositories or enemas.

Nasal Administration

Pharmaceutical compositions adapted for nasal administration may usesolid carriers, e.g., powders (e.g., having a particle size in the rangeof 20 to 500 microns). Powders can be administered in the manner inwhich snuff is taken, e.g., by rapid inhalation through the nose from acontainer of powder held close to the nose. Compositions adopted fornasal administration may alternatively use liquid carriers, e.g., nasalsprays or nasal drops. These may comprise aqueous or oil solutions ofthe active ingredient. Compositions for administration by inhalation maybe supplied in specially adapted devices, e.g., in pressurized aerosols,nebulizers, or insufflators. These devices can be constructed so as toprovide predetermined dosages of the active ingredient

Vaginal Administration

Pharmaceutical compositions adapted for vaginal administration may beprovided as pessaries, tampons, creams, gels, pastes, foams or sprayformulations.

Parenteral Administration

If the agent of the present technology is administered parenterally,then examples of such administration include one or more of:intravenously, intra-arterially, intraperitoneally, intrathecally,intraventricularly, intraurethrally, intrasternally, intracranially,intramuscularly or subcutaneously administering the agent; and/or byusing infusion techniques.

For parenteral administration, the agent is best used in the form of asterile aqueous solution which may contain other substances, forexample, enough salts or glucose to make the solution isotonic withblood. The aqueous solutions should be suitably buffered (preferably toa pH of from 3 to 9), if necessary. The preparation of suitableparenteral formulations under sterile conditions is readily accomplishedby standard pharmaceutical techniques well-known to those skilled in theart.

Transdermal; Transmucosal; Transurethral or Intraurethral

“Transdermal” refers to the delivery of a compound by passage throughthe skin and into the blood stream. “Transmucosal” refers to delivery ofa compound by passage of the compound through the mucosal tissue andinto the blood stream. “Transurethral” or “intraurethral” refers todelivery of a drug into the urethra, such that the drug contacts andpasses through the wall of the urethra and enters into the blood stream.

Penetration Enhancement or Permeation Enhancement

“Penetration enhancement” or “permeation enhancement” refers to anincrease in the permeability of the skin or mucosal tissue to a selectedpharmacologically active compound such that the rate at which thecompound permeates through the skin or mucosal tissue is increased.

Penetration enhancers may include, for example, dimethylsulfoxide(DMSO); dimethyl formamide (DMF); N,N-dimethylacetamide (DMA);decylmethylsulfoxide (CIOMSO); polyethyleneglycol monolaurate (PEGML);glyceral monolaurate; lecithin; 1-substituted azacycloheptanones,particularly 1-N-dodecylcyclaza-cycloheptanones (e.g., as availableunder the trademark Azone™ from Nelson Research & Development Co.,Irvine, Calif.), alcohols, and the like.

Carriers or Vehicles

“Carriers” or “vehicles” refers to carrier materials suitable forcompound administration and include any such material known in the artsuch as, for example, any liquid, gel, solvent, liquid diluent,solubilizer, or the like, which is non-toxic and which does not interactwith any components of the composition in a deleterious manner.

Examples of pharmaceutically acceptable carriers include, for example,water, salt solutions, alcohol, silicone, waxes, petroleum jelly,vegetable oils, polyethylene glycols, propylene glycol, sugars, gelatin,lactose, amylose, magnesium stearate, talc, surfactants, silicic acid,viscous paraffin, perfume oil, fatty acid monoglycerides anddiglycerides, petroethral fatty add esters, hydroxymethyl-cellulose,polyvinylpyrrolidone, and the like.

Epidermal Drug Delivery (Transfersomes)

Transfersomes (“carrying bodies”) are complex, most often vesicular, bi-or multi-component aggregates capable of crossing barriers and oftransferring material between the application and the destination sites.Transfersomes are sold by IDEA Corporation, Munich, Germany, andTRANSFERSOME is a trade mark of that company. Transfersome transdermaldrug delivery technology may be used for controllable and non-invasivedelivery of a wide variety of large molecules as well as for theimproved delivery of small molecules, including the metabolic enzymeantagonists and/or drugs of the present technology.

Transfersomes may be optimized to attain extremely flexible andself-regulating membranes. They are therefore deformable andconsequently can cross microporous barriers efficiently, even when theavailable passages are much smaller than the average aggregate size.Transfersome formulations are typically composed of natural amphipaticcompounds suspended in a water-based solution, optionally containingbiocompatible surfactants. Vesicular Transfersomes consist of a lipidbilayer surrounding an aqueous core and further contain at least onecomponent, capable of softening the membrane. The bilayer of aTransferosome is therefore more flexible than a liposome membrane, evenmetastable. Transfersome vesicles consequently change their shape easilyby adjusting locally to ambient stress.

Skin is one of the best biological barriers. Its outermost part reachesless than 10% into the depth of the skin but contributes over 80% to theskin permeability barrier. This body protecting layer consists ofoverlapping, flaccid corneocytes, organized in columnar clusters, sealedwith multilamellar lipid sheets that are covalently attached to the cellmembranes and very tightly packed. Generally, the average number of, andthe degree of order in, the intercellular lipid lamellae increasestoward the skin surface. This is accompanied by a continuous, butnonlinear, decrease in local water content near the surface.Notwithstanding this, the peak skin barrier is located in the inner halfof the outermost layer, where the intercellular lipid seals are alreadyformed, but not yet compromised by the skin cells detachment.

Passage of transfersome aggregates across the skin is a function ofvesicle membrane flexibility, hydrophilicity, and the ability to retainvesicle integrity, while the aggregate undergoes a significant change inshape. When a suspension of Transfersome vesicles is placed on thesurface of the skin, water evaporates from the relatively arid skinsurface and the vesicles start to dry out. Due to the strong polarity ofmajor Transfersome ingredients, the large number of hydrophilic groupson the membrane, assisted by the softness of the membrane, the vesiclesare attracted to the areas of higher water content in the narrow gapsbetween adjoining cells in the skin barrier, enabling skin penetrationof the vehicle. This, together with the vesicle's extreme ability todeform, enables Transfersome aggregates to open, temporarily, the tiny“cracks” through which water normally evaporates out of the skinChannels between the skin cells, two orders of magnitude wider than theoriginal micropores, are thus created. Such newly activated passages canaccommodate sufficiently deformable vesicles, which maintain theirintegrity but change their shape to fit the channel Along the resulting“virtual pathways”, or “virtual channels” in the outermost layer,Transfersomes reach regions of high water content in the deeper skinlayers. There, the vesicles (re)distribute. Since Transfersomes are toolarge to enter the blood vessels locally, they bypass the capillary bedand get to subcutaneous tissue, where they accumulate.

Although small molecules that have crossed the outermost layer of theskin (stratum corneum) are normally cleared from the skin through theblood circulation, delivery of drugs by means of Transfersome vesiclesallows accumulation of drug deep under the skin. Due to their largesize, the vesicles are cleared slowly from the skin and associated drugscan accumulate at the site. Transfersome mediated administration ofweight drugs, consequently, tends to shift the drug distribution towardsthe deep tissue under the application site.

Blood Brain Barrier (BBB)

Pharmaceutical compositions may be designed to pass across the bloodbrain barrier (BBB). For example, a carrier such as a fatty acid,inositol or cholesterol may be selected that is able to penetrate theBBB. The carrier may be a substance that enters the brain through aspecific transport system in brain endothelial cells, such asinsulin-like growth factor I or II. The carrier may be coupled to theactive agent or may contain and/or be in admixture with the activeagent. Liposomes can be used to cross the BBB. WO91/04014 describes aliposome delivery system in which an active agent can beencapsulated/embedded and in which molecules that are normallytransported across the BBB (e.g., insulin or insulin-like growth factorI or II) are present on the liposome outer surface. Liposome deliverysystems are also discussed in U.S. Pat. No. 4,704,355.

Polymer Delivery/Therapeutics

The agents may further be delivered attached to polymers. Polymer basedtherapeutics have been proposed to be effective delivery systems, andgenerally comprise one or more agents to be delivered attached to apolymeric molecule, which acts as a carrier. The agents are thusdisposed on the polymer backbone, and are carried into the target celltogether with the polymer.

The agents may be coupled, fused, mixed, combined, or otherwise joinedto a polymer. The coupling, etc. between the agent and the polymer maybe permanent or transient, and may involve covalent or non-covalentinteractions (including ionic interactions, hydrophobic forces, Van derWaals interactions, etc.). The exact mode of coupling is not importantas long as the agent is taken into a target cell substantially togetherwith the polymer. For simplicity, the entity comprising the agentattached to the polymer carrier is referred to here as a “polymer-agentconjugate”.

Any suitable polymer, for example, a natural or synthetic polymer, maybe used, e.g., the carrier polymer is a synthetic polymer such as PEG.In some embodiments, the carrier polymer is a biologically inertmolecule. Particular examples of polymers include polyethylene glycol(PEG), N-(2-hydroxypropyl) methacrylamide (HPMA) copolymers,polyamidoamine (PAMAM) dendrimers, HEMA, linear polyamidoamine polymers,etc. Any suitable linker for attaching the agent to the polymer may beused. In some embodiments, the linker is a biodegradable linker. Use ofbiodegradable linkers enables controlled release of the agent onexposure to the extracellular or intracellular environment. Highmolecular weight macromolecules are unable to diffuse passively intocells and are instead engulfed as membrane-encircled vesicles. Onceinside the vesicle, intracellular enzymes may act on the polymer-agentconjugate to effect release of the agent. Controlled intracellularrelease circumvents the toxic side effects associated with many drugs.

Furthermore, agents may be conjugated, attached, etc. by methods knownin the art to any suitable polymer and delivered. The agents may inparticular comprise any of the molecules referred to as “second agents”,such as polypeptides, nucleic acids, macromolecules, etc., as describedin the section below. In particular, the agent may comprise a pro-drugas described elsewhere.

The ability to choose the starting polymer enables the engineering ofpolymer-agent conjugates for desirable properties. The molecular weightof the polymer (and thus the polymer-agent conjugate), as well as itscharge and hydrophobicity properties, may be precisely tailored.Advantages of using polymer-agent conjugates include economy ofmanufacture, stability (longer shelf life), and reduction ofimmunogenicity and side effects. Furthermore, polymer-agent conjugatesare especially useful for the targeting of tumor cells because of theenhanced permeability and retention (EPR) effect, in which growingtumors are more “leaky” to circulating macromolecules and largeparticules, allowing them easy access to the interior of the tumor.Increased accumulation and low toxicity (typically 10-20% of thetoxicity of the free agent) are also observed. Use of hyperbrancheddendrimers, for example, PAMAM dendrimers, is particularly advantageousin that they enable monodisperse compositions to be made and alsoflexibility of attachment sites (within the interior or the exterior ofthe dendrimer). The pH responsiveness of polymer-agent conjugates, forexample, those conjugated to polyamindoamine polymers, may be tailoredfor particular intracellular environments. This enables the drug to bereleased only when the polymer therapeutic encounters a particular pH orrange of pH, e.g., within a particular intracellular compartment. Thepolymer agent conjugates may further comprise a targeting means, such asan immunoglobulin or antibody, which directs the polymer-agent conjugateto certain tissues, organs or cells comprising a target, for example, aparticular antigen. Other targeting means are described elsewhere inthis document, and are also known in the art.

Particular examples of polymer-agent conjugates include “Smancs”,comprising a conjugate of styrene-co-maleic anhydride and the antitumourprotein neocarzinostatin, and a conjugate of PEG (polyethylene glycol)with L-asparaginase for treatment of leukaemia; PK1 (a conjugate of aHPMA copolymer with the anticancer drug doxorubicin); PK2 (similar toPK1, but furthermore including a galactose group for targeting primaryand secondary liver cancer); a conjugate of HPMA copolymer with theanticancer agent captothecin; a conjugate of HPMA copolymer with theanticancer agent paclitaxel; HPMA copolymer-platinate, etc. Any of thesepolymer-agent conjugates are suitable for co-loading into the transgeniccells of the present technology.

Dose Levels

Typically, a physician will determine the actual dosage which will bemost suitable for an individual subject. The specific dose level andfrequency of dosage for any particular patient may be varied and willdepend upon a variety of factors including the activity of the specificcompound employed; the metabolic stability and length of action of thatcompound; the age, body weight, general health, sex, diet, mode and timeof administration; rate of excretion; drug combination; the severity ofthe particular condition; and the individual undergoing therapy. Theagent and/or the pharmaceutical composition of the present technologymay be administered in accordance with a regimen of from 1 to 10 timesper day, such as once or twice per day. For oral and parenteraladministration to human patients, the daily dosage level of the agentmay be in single or divided doses.

Depending upon the need, the agent may be administered at a dose of from0.01 to 30 mg/kg body weight, such as from 0.1 to 10 mg/kg or from 0.1to 1 mg/kg body weight. Naturally, the dosages mentioned herein areexemplary of the average case. There can, of course, be individualinstances where higher or lower dosage ranges are merited.

Therapeutically Effective Amount

“Therapeutically effective amount” refers to the amount of thetherapeutic agent that is effective to achieve its intended purpose.While individual patient needs may vary, determination of optimal rangesfor effective amounts of the compounds related to the technology iswithin the skill of the art. Generally, the dosage regimen for treatinga condition with the compounds and/or compositions of this technology isselected in accordance with a variety of factors, including the type,age, weight, sex, diet and medical condition of the patient; theseverity of the dysfunction; the route of administration;pharmacological considerations such as the activity, efficacy,pharmacokinetic and toxicology profiles of the particular compound used;whether a drug delivery system is used; and whether the compound isadministered as part of a drug combination and can be adjusted by oneskilled in the art. Thus, the dosage regimen actually employed may varywidely and therefore may deviate from the exemplary dosage regimens setforth herein.

Pharmaceutical Combinations

In general, the agent may be used in combination with one or more otherpharmaceutically active agents. Other agents are sometimes referred toauxiliary agents.

Pharmaceutically Acceptable Salt

The agent may be in the form of, and/or may be administered as, apharmaceutically acceptable salt, e.g., an acid addition salt or a basesalt, or a solvate thereof, including a hydrate thereof. For a review onsuitable salts see Berge et al, J. (1977) Pharm. Sci. 66: 1-19.

Typically, a pharmaceutically acceptable salt may be readily prepared byusing a desired acid or base, as appropriate. The salt may precipitatefrom solution and be collected by filtration or may be recovered byevaporation of the solvent.

Suitable acid addition salts are formed from acids that form non-toxicsalts such as hydrochloride, hydrobromide, hydroiodide, sulphate,bisulphate, nitrate, phosphate, hydrogen phosphate, acetate, maleate,fumarate, lactate, tartrate, citrate, gluconate, succinate, saccharate,benzoate, methanesulphonate, ethanesulphonate, benzenesulphonate,p-toluenesulphonate, and pamoate salts.

Suitable base salts are formed from bases that form non-toxic salts andexamples are sodium, potassium, aluminum, calcium, magnesium, zinc, anddiethanolamine salts.

Disease States

The present technology relates to the use of a composition according toembodiments of the technology for the manufacture of a medicament forthe treatment and/or prevention of a condition selected from diabetes,inflammatory disorders, metabolic syndrome, obesity, hypertension, fattyliver, diabetes, hyperglycaemia, hyperinsulinemia, and stenosis.

In some embodiments, the present technology provides use of a compoundaccording to the technology for the manufacture of a medicament forlowering concentration of cholesterol and triglycerides in the blood ofmammals and/or inhibiting the oxidative modification of low densitylipoprotein.

In some embodiments, the present technology provides a method forproducing weigh loss or a reduction of the fat mass in a human ornon-human animal in need thereof, comprising administering thereto aneffective amount of a compound of the technology or a pharmaceuticallyacceptable salt thereof.

In some embodiments, the present technology provides a method for themodification of the fat distribution and content of animals in order toimprove the quality of the meat, or product such as milk and eggs,comprising administering thereto an effective amount of a compound ofthe technology or a pharmaceutically acceptable salt thereof. Preferablysaid animal is an agricultural animal, such as gallinaceous birds,bovine, ovine, caprine or porcine mammals. The animal may be a fish orshellfish, such as salmon, cod, Tilapia, clams, oysters, lobster orcrabs.

In some embodiments, the present technology provides use of a compoundaccording to the technology or a pharmaceutically acceptable saltthereof in the manufacture of a medicament for the inhibition and/orprevention of the growth of tumors.

In some embodiments, the present technology provides use of a compoundaccording to the technology in the manufacture of a medicament for theinhibition and/or prevention of the invasion of a primary tumor into theconnective tissue.

In some embodiments, the present technology provides use of a compoundaccording to the technology for the manufacture of a medicament for theinhibition and/or prevention of the metastatic properties of a tumor,e.g., to inhibit the formation of secondary tumors. For example, the useof the present compounds may increase the overall survival of mammalswith tumors.

In some embodiments, the present technology provides a method for thetreatment and/or inhibition of primary and secondary metastaticneoplasms, comprising administering a compound of the technology or apharmaceutically acceptable salt thereof.

In some embodiments, the present technology provides use of a compoundof the technology or a pharmaceutically acceptable salt thereof in themanufacture of a medicament for the prevention and/or treatment ofproliferative skin disorders such as psoriasis, atopic dermatitis,non-specific dermatitis, primary irritant contact dermatitis, allergiccontact dermatitis, lamellar ichthyosis, epidermolytic hyperkeratosis,pre malignant sun induced keratosis, and seborrhea.

In some embodiments, the present technology provides use of a compoundof the technology or a pharmaceutically acceptable salt thereof in themanufacture of a medicament for the inhibition of proliferation and/orinduction of differentiation of keratinocytes.

In some embodiments, the present technology provides use of a compoundof the technology or a pharmaceutically acceptable salt thereof in themanufacture of a medicament for the prevention and/or treatment ofinflammatory disorders. For example, in some embodiments, the presenttechnology provides use of a compound of the technology or apharmaceutically acceptable salt thereof in the manufacture of amedicament for the prevention and/or treatment of inflammatorydisorders, wherein the inflammatory disorder is selected from the groupcomprising immune mediated disorders such as rheumatoid arthritis,systemic vasculitis, systemic lupus erythematosus, systemic sclerosis,dermatomyositis, polymyositis, various autoimmune endocrine disorders(e.g. thyroiditis and adrenalitis), various immune mediated neurologicaldisorders (e.g. multiple sclerosis and myastenia gravis), variouscardiovascular disorders (e.g. myocarditis, congestive heart failure,arteriosclerosis and stable and unstable angina, and Wegenersgranulomatosis), inflammatory bowel diseases and colitis (e.g., Crohn'scolitis), nephritis, various inflammatory skin disorders (e.g.psoriasis, atopic dermatitis and food allergy) and acute and chronicallograft rejection after organ transplantation.

In some embodiments, the present technology provides a method forenhancing the endogenous production of interleukin-10 (IL-10) inmammalian cells or tissues, comprising administering a compound of thetechnology or a pharmaceutically acceptable salt thereof. In someembodiments, the mammal has developed or is susceptible to develop anautoimmune and/or inflammatory disorder.

In some embodiments, the present technology provides a method forsuppression of the endogenous production of interleukin-2 (IL-2) inmammalian cells or tissues, comprising administering a compound of thetechnology or a pharmaceutically acceptable salt thereof. In someembodiments, the mammal has developed or is susceptible to develop anautoimmune and/or inflammatory disorder.

In some embodiments, the present technology provides use of a compoundof the technology or a pharmaceutically acceptable salt thereof in themanufacture of a medicament for the inhibition of proliferation ofstimulated peripheral mononuclear cells (PBMC).

Further description of these and other diseases is provided below.

Obesity and Related Diseases

Obesity is a chronic disease that is highly prevalent in modern societyand is associated not only with a social stigma, but also with decreasedlife span and numerous medical problems, including adverse psychologicaldevelopment, reproductive disorders such as polycystic ovarian disease,dermatological disorders such as infections, varicose veins, Acanthosisnigricans, and eczema, exercise intolerance, diabetes mellitus, insulinresistance, hypertension, hypercholesterolemia, cholelithiasis,osteoarthritis, orthopedic injury, thromboembolic disease, cancer, andcoronary heart disease.

In some embodiments, the present technology provides a treatment regimenthat is useful in returning the body weight of obese subjects toward anormal body weight. In some embodiments, the technology provides atherapy for obesity that results in maintenance of the lowered bodyweight for an extended period of time. Further, in some embodiments thepresent technology reduces or inhibits the weight gain normally inducedby fat rich diets.

In some embodiments, the present technology prevents obesity and, oncetreatment has begun, to arrests progression or prevents the onset ofdiseases that are the consequence of, or secondary to, the obesity, suchas hypertension and fatty liver.

The obesity herein may be due to any cause, whether genetic orenvironmental. Examples of disorders that may result in obesity or bethe cause of obesity include overeating and bulimia, polycystic ovariandisease, craniopharyngioma, the Prader-Willi Syndrome, Frohlich'ssyndrome, Type II diabetes, GH-deficiency, normal variant short stature,Turner's syndrome, and other pathological conditions showing reducedmetabolic activity.

In some embodiments, the present technology provides a treatment regimenthat is useful in lowering the blood pressure. Further, in someembodiments the present technology provides a treatment regimen that isuseful in lowering the concentration of triacylglycerols in the liver.It is anticipated that such a regimen provides an inhibiting effect onthe development of a fatty liver condition and is suited as a method forthe treatment of the manifested disease.

In some embodiments, the compounds of the present technology activatethe oxidation, and also reduce the concentration, of triglycerides inthe liver.

The term “metabolic syndrome” is used to describe a multimetabolicsyndrome that is inter alia characterized by hyperinsulinemia, insulinresistance, obesity, glucose intolerance, Type 2 diabetes mellitus,dyslipidemia, or hypertension.

As indicated above it is anticipated that the compounds of the presenttechnology provide a positive effect on all the conditions mentionedabove, e.g., by regulating both glucose and lipid homeostasis, and thusit is anticipated that the compounds of the present technology aresuitable agents for the regulation of the above defined metabolicdisease (sometimes called syndrome X).

Diabetes

There are two major forms of diabetes mellitus. One is type I diabetes,which is also known as insulin-dependent diabetes mellitus (IDDM), andthe other is type II diabetes, which is also known asnoninsulin-dependent diabetes mellitus (NIDDM). Most patients with IDDMhave a common pathological picture; the nearly total disappearance ofinsulin-producing pancreatic beta cells which results in hyperglycemia.

Considerable evidence has been accumulated showing that most IDDM is theconsequence of progressive beta-cell destruction during an asymptomaticperiod often extending over many years. The prediabetic period isrecognized usually by the detection of circulating islet-cellautoantibodies and insulin autoantibodies.

As such, there is a need for a compound that is nontoxic and has no orminimal side effects but that would prevent clinical IDDM and NIDDM.

Type I diabetes: severe diabetes mellitus, usually of abrupt onset priorto maturity, characterized by low plasma insulin levels, polydipsia,polyuria, increased appetite, weight loss and episodic ketoacidosis;also referred to as IDDM.

Type II diabetes: an often mild form of diabetes mellitus, often ofgradual onset, usually in adults, characterized by normal to highabsolute plasma insulin levels which are relatively low in relation toplasma glucose levels; also referred to as NIDDM.

Type I and II diabetes are in accordance with an etiologicclassification considered as primary diabetes respectively.

Secondary diabetes comprises pancreatic, extrapancreatic and/orendocrine or drug-induced diabetes. Further, some types of diabetes areclassified as exceptional forms. These include lipoatrophic, myatonicdiabetes, and a type of diabetes caused by disturbance of insulinreceptors.

Considering the high prevalence of diabetes in our society and theserious consequences associated therewith as discussed above, anytherapeutic drug potentially useful for the treatment and prevention ofthis disease would have a profound beneficial effect on their health.There is a need in the art for a drug that reduces the concentration ofglucose in the blood of diabetic subjects without significant adverseside effects.

Accordingly, in some embodiments, the present technology provides atreatment regimen that is useful in lowering the blood glucose and totreat a diabetic condition. Moreover, in some embodiments, the presenttechnology provides a treatment regimen that is useful in lowering theconcentration of insulin in the blood, and to increase the effect of theremaining insulin. In some preferred embodiments, the compositions ofthe present invention are useful for ameliorating the symptoms ofdiabetes, providing nutritional support to a subject with diabetes,promoting healthy blood sugar levels, supporting efficient insulinproduction and secretion, and/or supporting healthy glucose metabolism.

Stenosis

Many pathological conditions have been found to be associated withsmooth muscle cell proliferation. Such conditions include restenosis,arteriosclerosis, coronary heart disease, thrombosis, myocardialinfarction, stroke, smooth muscle neoplasms such as leiomyoma, andleiomyosarcoma of the bowel and uterus and uterine fibroid or fibroma.

Over half a million interventional intravascular procedures areperformed each year. While such invasive procedures continue to improveover time, as many as 30% to 50% of the procedures performed each yearfail as a result of restenosis, e.g., the formation of secondarystenosis. The reduction of restenosis is, therefore, often cited as themost critical factor in increasing the success realized in the treatmentof cardiovascular disease through the use of interventionalintravascular procedures, such as angioplasty, atherectomy, andprocedures utilizing stents, and laser technology.

In balloon angioplasty, e.g. Percutaneous Transluminal CoronaryAngioplasty (PTCA), a small incision is made to an artery in thepatient's leg or arm and a long hollow tube, called a guide catheter, isinserted into the artery. A thick guide wire and deflated ballooncatheter are then inserted into the guide catheter and are carefullyadvanced through the patient's blood vessels using X-ray visualization.The deflated balloon is advanced until it reaches the site of theluminal narrowing, at which point the physician inflates the balloon oneor more times to a pressure of about 4-6 atm for about 60 seconds. Wheninflated, the balloon cracks and fractures the plaque and stretches themuscle fiber in the artery wall beyond its ability to recoil completely.Although no plaque is removed in this procedure, the fracturing of theplaque and the stretching of the arterial wall increase the vessellumen, thereby allowing for increased blood flow.

The restenosis that accompanies such procedures is characterized byplatelet aggregation and adhesion, smooth muscle cell proliferation,narrowing of the vessel lumen, restricted vasodilatation, and anincrease in blood pressure. Smooth muscle cells in the intimal layer ofthe artery have been reported to enter the growth cycle within about 2-3days of these procedures and to proliferate for several days thereafter(intimal hyperplasia).

Compounds that reportedly suppress smooth muscle proliferation in vitromay have undesirable pharmacological side effects when used in vivo.Heparin is an example of one such compound, which reportedly inhibitssmooth muscle cell proliferation in vitro but when used in vivo has thepotential adverse side effect of inhibiting coagulation.

As is apparent from the foregoing, many problems remain to be solved inthe use of inhibitory drugs to effectively treat smooth muscle cellmobilization and proliferation. It would be highly advantageous todevelop new compositions or methods for inhibiting stenosis, restenosisor related disorders due to proliferation and mobilization of vascularsmooth muscle cells following, for example, traumatic injury to vesselsrendered during vascular surgery.

Accordingly, it is anticipated that embodiments of compounds inaccordance with the present technology are effective in the treatment ofthese diseases.

Tumors

The development of new and more effective chemotherapeutic agents forcancer treatment requires considering a variety of factors includingcytotoxicity, tumor cell proliferation, invasion, and metastasis.Conventional anticancer agents have typically been identified on thebasis of their cytotoxicity alone.

Tumor progression is thought to occur when variant cells havingselective growth properties arise within a tumor cell population, andone of the final stages of tumor progression is the appearance of themetastatic phenotype.

During metastasis, the tumor cells invade the blood vessels, surviveagainst circulating host immune defenses, and then extravasate, implant,and grow at sites distant from the primary tumor. This ability of tumorcells to invade neighboring tissues and to colonize other organs isamong the leading causes of cancer related deaths.

The term metastasis encompasses a number of phenotypic traits thattogether result in the clinical problem that most often leads to deathfrom cancer. The cells lose their adherence and restrained positionwithin an organized tissue, move into adjacent sites, develop thecapacity both to invade and to egress from blood vessels, and becomecapable of proliferating in unnatural locations or environments. Thesechanges in growth patterns are accompanied by an accumulation ofbiochemical alterations that have the capacity to promote the metastaticprocess.

So far, little is known about the intrinsic mechanism involved in themetastatic cascade. It is likely that in some cases the augmentedmetastatic potential of certain tumor cells may be due to an increasedexpression of oncogenes, which normally are responsible for control ofvarious cellular functions, including differentiation, proliferation,cell motility, and communication. Further, it has been shown thatsubstances that modulate signal transduction pathways can inhibit themetastatic behavior of a tumor, and it is also speculated that compoundswith surface related effects, e.g., compounds that modulates the cellmembranes, might be involved in the process leading to metastasis.

Cancer is a disease of inappropriate tissue accumulation. Thisderangement is most evident clinically when tumor tissue bulkcompromises the function of vital organs. Contrary to what is generallythought, human malignant disorders are usually not diseases of rapidcell proliferation. In fact, the cells of most common cancersproliferate more slowly than many cells in normal tissues. It is arelatively slow accumulation of tumor tissue within vital organs thatproves fatal to most patients who die of cancer.

Chemotherapeutic agents share one characteristic: they are usually moreeffective in killing or damaging malignant cells than normal cells.However, the fact that they do harm normal cells indicates theirpotential for toxicity. Nearly all chemotherapeutic agents currently inuse interfere with DNA synthesis, with the provision of precursors forDNA and RNA synthesis, or with mitosis. Such drugs are most effectiveagainst cycling cells. The mechanism of cell death after treatment withany single agent or combination of agents is complex and is likely toinclude more than one process. Because most clinically detectable tumorsare composed mostly of non-cycling cells, it is not surprising thatchemotherapy is not always effective in eradicating cancer.

The strategy of cancer treatment is to shift tumor cells from anon-cycling compartment to a cycling compartment. Several methods thatpromote this shift form the basis for combined-modality treatment.Surgery is most commonly used to reduce tumor size and thus facilitatere-entry of cancer cells into the cell cycle. After the primary tumor iscompletely removed, microscopic metastases may remain at distant sites.Because of their small size, the micrometastases are composedprincipally of cycling cells Small numbers of cells that remain atprimary tumor site are also likely to re-enter the cell cycle. Thus, theremaining cancer cells are often susceptible to chemotherapy. Radiationtherapy or chemotherapy alone can also be used to reduce tumor bulk andthus recruit cells into the cycling cell compartment.

Combination drug therapy is, therefore, the basis for most chemotherapyemployed at present. Combination chemotherapy uses the differentmechanisms of action and cytotoxic potentials of multiple drugs.However, even though the chemotherapeutic agents are more effective inkilling or damaging malignant cells than normal cells, the fact thatthey do harm normal cells indicates their great potential for toxicity.For chemotherapy to be effective, the patient must be in goodphysiologic condition.

Cancer treatment requires inhibition of a variety of factors includingtumor cell proliferation, metastatic dissemination of cancer cells toother parts of the body, invasion, tumor-induced neovascularization, andenhancement of host immunological responses and cytotoxity.

Conventional cancer chemotherapeutic agents have often been selected onthe basis of their cytotoxicity to tumor cells. However, some anticanceragents have adverse effects on the patient's immunological system.Unfortunately, for the vast majority of conventional antineoplasticagents the margin between an effective dose and a toxic dose, e.g., thetherapeutic index, is extremely low. Thus, it would be greatlyadvantageous if a cancer therapy or treatment could be developed thatwould afford noncytotoxic protection against factors that might lead togrowth, progression and metastasis of invasive cancers.

Accordingly, in some embodiments, the present technology provides amethod for the prevention and/or treatment of primary and metastaticneoplasms that involves using a fatty acid analogue, or a lipidcomprising a fatty acid analogue, of the present technology to treat apatient suffering from a cancer.

The two essential features of cancer are invasion and metastasis. At oneextreme, microinvasion of the basement membrane characterizes thetransition from neoplasia to cancer, and at the other extreme,metastases generally lead to death. Invasion into the underlyingconnective tissue by primary tumor proceeds in stages and is facilitatedby various mediators produced by the tumor cells. Tumor cells that havenot invaded the basement membrane and remain confined within theepithelium are termed carcinoma in situ. Metastases, on the other hand,may form when circulating tumor cells with adherent lymphocytes andplatelets are trapped in capillaries and the tumor cell membraneinteracts with the capillary endothelium. The capillary endothelialjunctions retract, and tumor cell ligands bind to receptors on theendothelial and basement membranes.

Tumor cells then release collagenase IV, which destroys collagen IV, amajor component of the underlying basement membrane. Invasion of thesubcapillary connective tissue is aided by binding to the glycoproteinslaminin and fibronectin, by the release of proteases that destroy thematrix, and by the secretion of motility and chemotactic factors. Tumorcells then may proliferate and synthesise platelet aggregatory factorssuch as thromboxanes and procoagulants, thereby leading to thedeposition of a fibrin cocoon around the cells. Such a cocoon mayprotect the micrometastasis from attack by the host's immune system.

Cancers that can be prevented and/or treated by the compositions andmethods of the present technology include, but are not limited to, humansarcomas and carcinomas, e.g. carcinomas, e.g., colon carcinoma,pancreatic cancer, breast cancer, ovarian cancer, prostate cancer,thyroid cancer, fibrosarcoma, myxosarcoma, liposarcoma, chondrosarcoma,osteogenic sarcoma, chordoma, angiosarcoma, endotheliosarcoma,lymphangiosarcoma, lymphangioendotheliosarcoma, synovioma, mesothelioma,Ewing's tumor, leiomyosarcoma, rhabdomyosarcoma, squamous cellcarcinoma, basal cell carcinoma, adenocarcinoma, sweat gland carcinoma,sebaceous gland carcinoma, papillary carcinoma, papillaryadenocarcinomas, cystadenocarcinoma, medullary carcinoma, bronchogeniccarcinoma, renal cell carcinoma, hepatoma, bile duct carcinoma,choriocarcinoma, seminoma, embryonal carcinoma, Wilms's tumor, cervicalcancer, testicular tumor, lung carcinoma, small cell lung carcinoma,bladder carcinoma, epithelial carcinoma, glioma, astrocytoma,medulloblastoma, craniopharyngioma, ependymoma, pinealoma,hemangioblastoma, acoustic neuroma, oligodendroglioma, meningioma,melanoma, neuroblastoma, retinoblastoma, leukemias, e.g., acutelymphocytic leukemia and acute myelocytic leukemia (myeloblastic,promyelocytic, myelomonocytic, monocytic and erythroleukemia); chronicleukemia (chronic myelocytic (granulocytic) leukemia and chroniclymphocytic leukemia); and polycythemia vera, lymphoma (Hodgkin'sdisease and non-Hodgkin's disease), multiple myeloma, Waldenstrom'smacroglobulinemia, and heavy chain disease. Specific examples of suchcancers are described in the sections below.

Skin Disorders

Proliferative skin diseases are widespread throughout the world andafflict millions of humans and their domesticated animals. Proliferativeskin diseases are characterized by keratinocyte cell proliferation, ordivision, and may also be associated with incomplete epidermaldifferentiation. Psoriasis is the most serious of the proliferative skindiseases with which this technology is concerned.

Psoriasis is a genetically determined disease of the skin characterizedby two biological hallmarks. First, there is a profound epidermalhyperproliferation related to accelerated and incompletedifferentiation. Second, there is a marked inflammation of bothepidermis and dermis with an increased recruitment of T lymphocytes, andin some cases, formation of neutrophil microabcesses. Many pathologicfeatures of psoriasis can be attributed to alterations in the growth andmaturation of epidermal keratinocytes, with increased proliferation ofepidermal cells, occurring within 0.2 mm of the skin's surface.

Traditional investigations into the pathogenesis of psoriasis havefocused on the increased proliferation and hyperplasia of the epidermis.In normal skin, the time for a cell to move from the basal layer throughthe granular layer is 4 to 5 weeks. In psoriatic lesions, the time isdecreased sevenfold to tenfold because of a shortened cell cycle time,an increase in the absolute number of cells capable of proliferating,and an increased proportion of cells that are actually dividing. Thehyperproliferative phenomenon is also expressed, although to asubstantially smaller degree, in the clinically uninvolved skin ofpsoriatic patients.

A common form of psoriasis, psoriasis vulgaris, is characterized bywell-demarcated erythematous plaques covered by thick, silvery scales. Acharacteristic finding is the isomorphic response (Koebner phenomenon),in which new psoriatic lesions arise at sites of cutaneous trauma.Lesions are often localized to the extensor surfaces of the extremities,and the nails and scalp are also commonly involved.

Therapeutic efforts in psoriasis are aimed at decreasing theproliferative rate of the epidermis, either by direct action on celldivision or indirectly by reducing the immunological response. Forpatients with localized, limited psoriasis, administration of topicalcorticosteroids is the most convenient outpatient therapy.

Rapid improvement may be seen with this approach, but the beneficialshort-term efficacy is limited and chronic topical corticosteroidtreatment is not advisable. Side effects from chronic topicalcorticosteroid therapy can include atrophy of the skin, development oftolerance to the agent used (tachyphylaxis), and serious exacerbation ofthe disease after discontinuation. Pituitary-adrenal suppression is apotential and serious complication of potent topical corticosteroidtherapy, particularly when the agent covers a large portion of the bodysurface and is used under occlusive dressings.

The retinoids, particularly etretinate, either alone or in combinationwith PUVA, are also an effective treatment for psoriasis. Etretinate isespecially useful in the exfoliative and pustular varieties ofpsoriasis. However, several major potential complications must bemonitored in patients placed on retinoids. As a class, the retinoids arepotent teratogens and should not be given to women of childbearing agewho are not using adequate contraception.

Etretinate, like other retinoids, can produce elevations in cholesteroland triglyceride levels; therefore dietary regulation may be necessary.In addition, because etretinate can induce hepatotoxicity, liverfunction tests should be performed before and at regular intervalsduring use of the drug.

Considering the complications and side effects attendant to the use ofdifferent drugs and photochemotherapy currently used in treating a skinproliferative disease such as psoriasis, there is a need for a newmethod and a new composition to inhibit keratinocyte proliferation toalleviate the symptoms of skin proliferation diseases.

Inflammatory and Auto-Immune Disorders

Interleukins, interferons, colony stimulating factors and TNF-alpha areexamples of a group of diverse multi-functional proteins calledcytokines. Cytokines are a class of secreted soluble proteins normallypresent in very low concentration in a variety of cells. Lymphoid,inflammatory hemopoietic, and other cells such as connective tissuecells (e.g. fibroblasts, osteoblasts) secrete a variety of cytokineswhich regulate the immune, inflammatory, repair, and acute phaseresponses by controlling cell proliferation, differentiation, andeffector functions. The effects of cytokines are mediated throughbinding to high affinity receptors on specific cell types.

An important cytokine is IL-10, a 35-40 kDa peptide produced by helperT-cells, B-cells, monocytes, macrophages, and other cell types. Invitro, IL-10 has demonstrated immunosuppressive properties as evidencedby its ability to suppress cytokine production including IL-1 and TNFa.IL-10 also inhibits activation of other inflammatory cytokines, andtherefore has potent anti-inflammatory activity.

It has been of recent interest to administer-IL-10 in the treatment ofcertain conditions characterized by excessive IL-1 and TNF-alphaproduction. Such diseases or conditions include loosening of prostheticjoint implants, inflammation, diabetes, cancer, graft versus hostdiseases, viral, fungal and bacterial infections, lipopolysaccharideendotoxin shock, diseases of depressed bone marrow function,thrombocytopenia, osteoporosis, spondyloarthropathies, Paget's disease,inflammatory bowel disease, arthritis, osteoarthritis, autoimmunediseases such as rheumatoid arthritis, systemic lupus erythematosus, andconnective tissue diseases.

For example, purified IL-10 has been shown in vitro to suppress certaintypes of viral infections. U.S. Pat. No. 5,665,345 discloses a methodfor inhibiting replication of the human immunodeficiency virus,retro-viruses, and Kaposi sarcoma in human cells by administering IL-10.

IL-10 has also been suggested for use in the treatment of certaincancers. U.S. Pat. No. 5,570,190 discloses administering exogenous IL-10to treat mammals suffering from acute myelogenous leukemia and acutelymphocytic leukemia. IL-10 is said to be administered either in thepurified or recombinant form and is believed to inhibit theproliferation of acute leukemia blast cells. Similarly, IL-10 was shownto inhibit bone marrow metastasis in severe combined immunodeficientmice.

The above conventional approaches to treating conditions characterizedby excessive IL-1 and TNF-alpha production have been limited toadministering exogenous purified or recombinant IL-10 intravenously.Since IL-10 is a protein, it is difficult to infuse intravenously into amammal because proteins often leach out of solution and bind to theplastic or glass used in intravenous administration sets. Also, proteinsare often incompatible and precipitate when mixed with physiologicalsolutions such as dextrose or saline. In addition, oral and topicalroutes are unavailable for IL-10 administration. The oral route isunavailable because protein is degraded in the gastrointestinal tract.None of the above approaches suggests enhancing endogenous IL-10production in mammals for prophylaxis and treatment of diseases orconditions.

Further, it is known that IL-10 is a powerful deactivator of macrophagesand T cells, and inadequate production has been implicated in variousautoimmune and inflammatory disorders.

In addition, or in the alternative, embodiments of the compound orcomposition of the present technology are useful in the treatment of thefollowing disorders: cancer, inflammation or inflammatory disease,dermatological disorders, fever, cardiovascular effects, hemorrhage,coagulation and acute phase response, cachexia, anorexia, acuteinfection, HIV infection, shock states, graft-versus-host reactions,autoimmune disease, reperfusion injury, meningitis, migraine andaspirin-dependent anti-thrombosis; tumor growth, invasion and spread,angiogenesis, metastases, malignant, ascites and malignant pleuraleffusion; cerebral ischemia, ischaemic heart disease, osteoarthritis,rheumatoid arthritis, osteoporosis, asthma, multiple sclerosis, neurodegeneration, Alzheimer's disease, atherosclerosis, stroke, vasculitis,Crohn's disease and ulcerative colitis; periodontitis, gingivitis;psoriasis, atopic dermatitis, chronic ulcers, epidermolysis bullosa;corneal ulceration, retinopathy and surgical wound healing; rhinitis,allergic conjunctivitis, eczema, anaphylaxis; restenosis, congestiveheart failure, endometriosis, atherosclerosis or endosclerosis.

In addition, or in the alternative, embodiments of the compound orcomposition of the present technology are useful in the treatment of thefollowing disorders: cytokine and cell proliferation/differentiationactivity; immunosuppressant or immunostimulant activity (e.g., fortreating immune deficiency, including infection with human immunedeficiency virus; regulation of lymphocyte growth; treating cancer andmany autoimmune diseases, and to prevent transplant rejection or inducetumor immunity); regulation of hematopoiesis, e.g. treatment of myeloidor lymphoid diseases; promoting growth of bone, cartilage, tendon,ligament and nerve tissue, e.g. for healing wounds, treatment of burns,ulcers and periodontal disease and neurodegeneration; inhibition oractivation of follicle-stimulating hormone (modulation of fertility);chemotactic/chemokinetic activity (e.g. for mobilizing specific celltypes to sites of injury or infection); hemostatic and thrombolyticactivity (e.g. for treating hemophilia and stroke); anti-inflammatoryactivity (for treating e.g. septic shock or Crohn's disease); asantimicrobials; modulators of e.g. metabolism or behavior; asanalgesics; treating specific deficiency disorders; in treatment of e.g.psoriasis, in human or veterinary medicine.

In addition, or in the alternative, embodiments of the composition ofthe present technology are useful in the treatment of the followingdisorders: macrophage inhibitory and/or T cell inhibitory activity andthus, anti-inflammatory activity; anti-immune activity, e.g., inhibitoryeffects against a cellular and/or humoral immune response, including aresponse not associated with inflammation; inhibit the ability ofmacrophages and T cells to adhere to extracellular matrix components andfibronectin, as well as up-regulated fas receptor expression in T cells;inhibit unwanted immune reaction and inflammation including arthritis,including rheumatoid arthritis, inflammation associated withhypersensitivity, allergic reactions, asthma, systemic lupuserythematosus, collagen diseases and other autoimmune diseases,inflammation associated with atherosclerosis, arteriosclerosis,atherosclerotic heart disease, reperfusion injury, cardiac arrest,myocardial infarction, vascular inflammatory disorders, respiratorydistress syndrome or other cardiopulmonary diseases, inflammationassociated with peptic ulcer, ulcerative colitis and other diseases ofthe gastrointestinal tract, hepatic fibrosis, liver cirrhosis or otherhepatic diseases, thyroiditis or other glandular diseases,glomerulonephritis or other renal and urologic diseases, otitis or otheroto-rhino-laryngological diseases, dermatitis or other dermal diseases,periodontal diseases or other dental diseases, orchitis orepididimo-orchitis, infertility, orchidal trauma or other immune-relatedtesticular diseases, placental dysfunction, placental insufficiency,habitual abortion, eclampsia, pre-eclampsia and other immune and/orinflammatory-related gynaecological diseases, posterior uveitis,intermediate uveitis, anterior uveitis, conjunctivitis, chorioretinitis,uveoretinitis, optic neuritis, intraocular inflammation, e.g., retinitisor cystoid macular oedema, sympathetic ophthalmia, scleritis, retinitispigmentosa, immune and inflammatory components of degenerative fondusdisease, inflammatory components of ocular trauma, ocular inflammationcaused by infection, proliferative vitreo-retinopathies, acute ischaemicoptic neuropathy, excessive scarring, e.g. following glaucoma filtrationoperation, immune and/or inflammation reaction against ocular implantsand other immune and inflammatory-related ophthalmic diseases,inflammation associated with autoimmune diseases or conditions ordisorders where, both in the central nervous system (CNS) or in anyother organ, immune and/or inflammation suppression would be beneficial,Parkinson's disease, complication and/or side effects from treatment ofParkinson's disease, AIDS-related dementia complex HIV-relatedencephalopathy, Devic's disease, Sydenham chorea, Alzheimer's diseaseand other degenerative diseases, conditions or disorders of the CNS,inflammatory components of stokes, post-polio syndrome, immune andinflammatory components of psychiatric disorders, myelitis,encephalitis, subacute sclerosing pan-encephalitis, encephalomyelitis,acute neuropathy, subacute neuropathy, chronic neuropathy,Guillaim-Barre syndrome, Sydenham chora, myasthenia gravis, pseudo-tumorcerebri, Down's Syndrome, Huntington's disease, amyotrophic lateralsclerosis, inflammatory components of CNS compression or CNS trauma orinfections of the CNS, inflammatory components of muscular atrophies anddystrophies, and immune and inflammatory related diseases, conditions ordisorders of the central and peripheral nervous systems, post-traumaticinflammation, septic shock, infectious diseases, inflammatorycomplications or side effects of surgery, bone marrow transplantation orother transplantation complications and/or side effects, inflammatoryand/or immune complications and side effects of gene therapy, e.g. dueto infection with a viral carrier, or inflammation associated with AIDS,to suppress or inhibit a humoral and/or cellular immune response, totreat or ameliorate monocyte or leukocyte proliferative diseases, e.g.leukaemia, by reducing the amount of monocytes or lymphocytes, for theprevention and/or treatment of graft rejection in cases oftransplantation of natural or artificial cells, tissue and organs suchas cornea, bone marrow, organs, lenses, pacemakers, natural orartificial skin tissue.

Treatment

Embodiments of the technology include any therapeutic application thatcan benefit a human or non-human animal, for example a mammal. As such,both human and veterinary treatments are within the scope of the presenttechnology.

Treatment may be in respect of an existing condition or it may beprophylactic. It may be of an adult, a juvenile, an infant, a fetus, ora part of any of the aforesaid (e.g., an organ, tissue, cell, or nucleicacid molecule).

In some embodiments, an active agent for use in treatment isadministered via any appropriate route and at any appropriate dosage.Dosages can vary between wide limits, depending upon the nature of thetreatment, the age and condition of the individual to be treated, etc.,and a physician will ultimately determine appropriate dosages to beused. However, without being bound by any particular dosages, a dailydosage of a compound of the present technology of from 1 μg to 1 mg/kgbody weight may be suitable. The dosage may be repeated as often asappropriate. If side effects develop, the amount and/or frequency of thedosage can be reduced, in accordance with good clinical practice.

Polymorphic Form(s) and/or Asymmetric Carbon(s)

Embodiments of compounds according to the present technology may existin a polymorphic form. In addition, embodiments of compounds accordingto the present technology may contain one or more asymmetric carbonatoms and therefore exist in two or more stereoisomeric forms. Where anagent contains an alkenyl or alkenylene group, cis (E) and trans (Z)isomerism may also occur. The present technology includes the individualstereoisomers of the agent and, where appropriate, the individualtautomeric forms thereof, together with mixtures thereof.

Separation of diastereoisomers and/or cis and trans isomers may beachieved by conventional techniques, e.g., by fractionalcrystallisation, chromatography, or H.P.L.C. of a stereoisomeric mixtureof the agent or a suitable salt or derivative thereof. An individualenantiomer of a compound of the agent may also be prepared from acorresponding optically pure intermediate or by resolution, such as byH.P.L.C. of the corresponding racemate using a suitable chiral supportor by fractional crystallisation of the diastereoisomeric salts formedby reaction of the corresponding racemate with a suitable opticallyactive acid or base, as appropriate.

Isotopic Variations

The present technology also includes all suitable isotopic variations ofthe agent or a pharmaceutically acceptable salt thereof. An isotopicvariation of an agent of the present technology or a pharmaceuticallyacceptable salt thereof is defined as one in which at least one atom isreplaced by an atom having the same atomic number but an atomic massdifferent from the atomic mass usually found in nature. Examples ofisotopes that can be incorporated into the agent and pharmaceuticallyacceptable salts thereof include isotopes of hydrogen, carbon, nitrogen,oxygen, phosphorus, sulfur, fluorine and chlorine such as ²H, .³H, ¹³C,¹⁵N, ¹⁷O, ¹⁸O, ³¹P, ³²P, ³⁵S, ¹⁸F, and ³⁶Cl, respectively. Certainisotopic variations of the agent and pharmaceutically acceptable saltsthereof, for example, those in which a radioactive isotope such as ³H or¹⁴C is incorporated, are useful in drug and/or substrate tissuedistribution studies. Tritiated, e.g., ³H, and carbon-14, e.g., ¹⁴C,isotopes are particularly useful for their ease of preparation anddetectability. Further, substitution with isotopes such as deuterium,e.g., ²H, may afford certain therapeutic advantages resulting fromgreater metabolic stability, for example, increased in vivo half-life orreduced dosage requirements and hence may be preferred in somecircumstances. Isotopic variations of the agent of the presenttechnology and pharmaceutically acceptable salts thereof of thistechnology can generally be prepared by conventional procedures usingappropriate isotopic variations of suitable reagents.

Pro-Drug

In some embodiments, compounds according to the technology are derivedfrom a prodrug. Prodrugs are entities that may or may not possesspharmacological activity as such, but may be administered (such asorally or parenterally) and thereafter subject to bioactivation (forexample metabolized) in the body to form the agent of the presenttechnology which is pharmacologically active. Examples of prodrugsinclude entities that have certain protected group(s) and that may notpossess pharmacological activity as such, but may, in certain instances,be administered (such as orally or parenterally) and thereaftermetabolized in the body to form the agent of the present technology thatare pharmacologically active.

Pro-Moiety

In some embodiments, the technology encompasses certain moieties knownas “pro-moieties”, for example as described in “Design of Prodrugs” byH. Bundgaard, Elsevier, 1985 (the disclosure of which is herebyincorporated by reference). In some embodiments, a pro-moiety may beplaced on appropriate functionalities of the agents. Such prodrugs arealso included within the scope of the technology.

Derivative

The term “derivative” or “derivatized” as used herein includes chemicalmodification of an agent. Illustrative of such chemical modificationswould be replacement of hydrogen by a halo group, an alkyl group, anacyl group, or an amino group.

Chemical Modification

In one embodiment of the present technology, the agent may be achemically modified agent. The chemical modification of an agent of thepresent technology may either enhance or reduce hydrogen bondinginteraction, charge interaction, hydrophobic interaction, Van Der Waalsinteraction, or dipole interaction between the agent and the target. Insome embodiments, the identified agent may act as a model (for example,a template) for the development of other compounds.

Although the disclosure herein refers to certain illustratedembodiments, it is to be understood that these embodiments are presentedby way of example and not by way of limitation.

All publications and patents mentioned in the above specification areherein incorporated by reference in their entirety for all purposes.Various modifications and variations of the described compositions,methods, and uses of the technology will be apparent to those skilled inthe art without departing from the scope and spirit of the technology asdescribed. Although the technology has been described in connection withspecific exemplary embodiments, it should be understood that thetechnology as claimed should not be unduly limited to such specificembodiments. Indeed, various modifications of the described modes forcarrying out the technology that are obvious to those skilled inpharmacology, biochemistry, medical science, or related fields areintended to be within the scope of the following claims.

We claim:
 1. A method of treating a skin disorder selected form thegroup consisting of psoriasis, atopic dermatitis, non-specificdermatitis, primary irritant contact dermatitis, allergic contactdermatitis, lamellar ichthyosis, epidermolytic hyperkeratosis, premalignant sun induced keratosis, and seborrhea comprising contacting theskin of an individual in need thereof with a topical formulationcomprising a first lipid component comprising at least 10% w/w of a5,11,14-eicosatrienoic acid ethyl esters and a second lipid componentcomprising at least 1% w/w selected from the group consisting of aconjugated linoleic acid moiety and an omega-3 fatty acid moietyselected from the group consisting of anall-cis-5,8,11,14,17-eicosapentaenoic acid moiety, anall-cis-7,10,13,16,19-docosapentaenoic acid moiety, and anall-cis-4,7,10,13,16,19-docosahexaenoic acid moiety and combinationsthereof.
 2. The method of claim 1, wherein said at least one omega-3fatty acid moiety is selected from the group consisting free fattyacids, acylglycerides, phospholipids and esters comprising said at leastone omega-3 fatty acid moiety.
 3. The method of claim 1, wherein said atleast one conjugated linoleic acid moiety is selected from the groupconsisting free fatty acids, acylglycerides, phospholipids and esterscomprising said at least one conjugated linoleic acid moiety.
 4. Themethod of claim 1, wherein said first lipid component comprises at least20%, 30%, 40%, 50%, 60%, 70%, 80%, 90%, 95%, or 99% w/w of said5,11,14-eicosatrienoic acid moiety.
 5. The method of claim 1, whereinsaid composition comprises at least 5%, 10%, 20%, 30%, 40%, 50%, 60%,70%, 80%, 90%, 95%, or 99% w/w of said second lipid component.
 6. Themethod of claim 1, further comprising at least one pharmaceuticallyacceptable carrier.
 7. The method of claim 1, wherein said compositioncomprises an oil, powder, crystal, wax, emulsion, micelle, vesicle, orfilm.
 8. The method of claim 1, wherein said skin disorder is psoriasis.9. The method of claim 1, wherein said skin disorder is primary irritantcontact dermatitis.
 10. The method of claim 1, wherein said skindisorder is allergic contact dermatitis.
 11. The method of claim 1,wherein said skin disorder is lamellar ichthyosis.
 12. The method ofclaim 1, wherein said skin disorder is epidermolytic hyperkeratosis. 13.The method of claim 1, wherein said skin disorder is pre malignant suninduced keratosis.